Antiviral compounds

ABSTRACT

The disclosure is related to anti-viral compounds, compositions containing such compounds, and therapeutic methods that include the administration of such compounds, as well as to processes and intermediates useful for preparing such compounds.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a Continuation of Divisional U.S. application Ser.No. 15/679,014, filed Aug. 16, 2017. U.S. application Ser. No.15/246,074, filed Aug. 24, 2016 is a divisional of U.S. application Ser.No. 15/246,074, which is a continuation of U.S. application Ser. No.14/957,988, filed Dec. 3, 2015, which is a continuation of U.S.application Ser. No. 13/830,346, filed Mar. 14, 2013, and issued as U.S.Pat. No. 9,233,974 on Jan. 12, 2016, which claims the benefit under 35U.S.C. § 119(e) of U.S. Provisional Application Ser. No. 61/745,452,filed on Dec. 21, 2012, the entireties of which are incorporated hereinby reference.

BACKGROUND

Hepatitis C is recognized as a chronic viral disease of the liver whichis characterized by liver disease. Although drugs targeting the liverare in wide use and have shown effectiveness, toxicity and other sideeffects have limited their usefulness. Inhibitors of hepatitis C virus(HCV) are useful to limit the establishment and progression of infectionby HCV as well as in diagnostic assays for HCV.

There is a need for new HCV therapeutic agents. In particular, there isa need for HCV therapeutic agents that have broad activity against HCVgenotypes (e.g. genotypes 1a, 1b, 2a, 3a, 4a). There is also aparticular need for agents that are less susceptible to viralresistance. Resistance mutations to inhibitors have been described forHCV NS5A for genotypes 1a and 1b in Antimicrobial Agents andChemotherapy, September 2010, Volume 54, p. 3641-3650.

SUMMARY

The present disclosure provides compounds for use in pharmaceuticalcompositions and methods for treating hepatitis C (HCV). In particular,provided herein are compounds having a polycyclic core and at least one2,6-dimethyltetrahydro-2H-pyran-4-yl, 4-methyltetrahydro-2H-pyran-4-yl,or tetrahydro-2H-pyran-3-yl capping group, which compounds exhibitbeneficial properties, such as, for example, enhanced bioavailabilityand/or enhanced activity against certain HCV genotypes, including butnot limited to, known resistant mutations thereof (see, e.g., Tables 1,2A and 2B).

In one embodiment the disclosure provides a compound of formula (I):

E^(1a)-V^(1a)—C(═O)—P^(1a)—W^(1a)—P^(1b)—C(═O)—V^(1b)-E^(1b)  (I)

wherein:

W^(1a) is

and W^(1a) is optionally substituted with one or more halo, alkyl,haloalkyl, optionally substituted aryl, optionally substitutedheterocycle, or cyano;

Y⁵ is —O—CH₂—, —CH₂—O—, —O—C(═O)—, or —C(═O)—O—;

X⁵ is —CH₂—CH₂—, or —CH═CH—;

P^(1a) and P^(1b) are each independently:

V^(1a) and V^(1b) are each independently:

provided that at least one of V^(1a) and V^(1b) is

E^(1a) and E^(1b) are each independently —N(H)(alkoxycarbonyl),—N(H)(cycloalkylcarbonyl), or —N(H)(cycloalkyloxycarbonyl); orE^(1a)-V^(1a) taken together are R^(9a); or E^(1b)-V^(1b) taken togetherare R^(9b); and

R^(9a) and R^(9b) are each independently:

or a pharmaceutically acceptable salt or prodrug thereof.

The disclosure also provides isotopically enriched compounds that arecompounds of the disclosure that comprise an enriched isotope at one ormore positions in the compound.

The present disclosure also provides a pharmaceutical compositioncomprising a compound of the disclosure or a pharmaceutically acceptablesalt or prodrug thereof and at least one pharmaceutically acceptablecarrier.

The present disclosure also provides a pharmaceutical composition foruse in treating hepatitis C (HCV). In one embodiment the compositioncomprises at least one additional therapeutic agent for treating HCV. Inone embodiment, the therapeutic agent is selected from ribavirin, an NS3protease inhibitor, a nucleoside or nucleotide inhibitor of HCV NS5Bpolymerase, an alpha-glucosidase 1 inhibitor, a hepatoprotectant, anon-nucleoside inhibitor of HCV polymerase, or combinations thereof. Inone embodiment, the composition further comprises a nucleoside ornucleotide inhibitor of HCV NS5B polymerase. In one embodiment, thenucleoside or nucleotide inhibitor of HCV NS5B polymerase is selectedfrom ribavirin, viramidine, levovirin, a L-nucleoside, or isatoribine.

In one embodiment, provided is a pharmaceutical composition comprising acompound as described herein and at least one nucleoside or nucleotideinhibitor of HCV NS5B polymerase, and at least one pharmaceuticallyacceptable carrier. In one embodiment, the composition further comprisesan interferon, a pegylated interferon, ribavirin or combinationsthereof. In one embodiment, the nucleoside or nucleotide inhibitor ofHCV NS5B polymerase is sofosbuvir. In one embodiment, provided is apharmaceutical composition comprising a compound as described herein andat least one NS3 protease inhibitor, and at least one pharmaceuticallyacceptable carrier. In one embodiment, the composition further comprisessofosbuvir.

The present disclosure also provides a pharmaceutical compositionfurther comprising an interferon or pegylated interferon.

The present disclosure also provides a pharmaceutical compositionfurther comprising a nucleoside analog.

The present disclosure also provides for a pharmaceutical compositionwherein said nucleoside analogue is selected from ribavirin, viramidine,levovirin, an L-nucleoside, and isatoribine and said interferon isα-interferon or pegylated α-interferon.

The present disclosure also provides for a method of treating hepatitisC, said method comprising administering to a human patient apharmaceutical composition which comprises a therapeutically effectiveamount of a compound of the disclosure.

The present disclosure also provides a method of inhibiting HCV,comprising administering to a mammal afflicted with a conditionassociated with HCV activity, an amount of a compound of the disclosure,effective to inhibit HCV.

The present disclosure also provides a compound of the disclosure foruse in medical therapy (e.g. for use in inhibiting HCV activity ortreating a condition associated with HCV activity), as well as the useof a compound of the disclosure for the manufacture of a medicamentuseful for inhibiting HCV or the treatment of a condition associatedwith HCV activity in a mammal.

The present disclosure also provides synthetic processes and novelintermediates disclosed herein which are useful for preparing compoundsof the disclosure. Some of the compounds of the disclosure are useful toprepare other compounds of the disclosure.

In another aspect the disclosure provides a compound of the disclosure,or a pharmaceutically acceptable salt or prodrug thereof, for use in theprophylactic or therapeutic treatment of hepatitis C or a hepatitis Cassociated disorder.

In another aspect the disclosure provides a method of inhibiting HCVactivity in a sample comprising treating the sample with a compound ofthe disclosure.

Compounds of formula (I) have been found to possess useful activityagainst several HCV genotypes. Additionally certain compounds of formula(I) exhibit significant potency against resistant variants in, e.g.,GT1.

DETAILED DESCRIPTION

Reference will now be made in detail to certain embodiments of thedisclosure, examples of which are illustrated in the accompanyingstructures and formulas. While the disclosure will be described inconjunction with the enumerated embodiments, it will be understood thatthey are not intended to limit the disclosure to those embodiments. Onthe contrary, the disclosure is intended to cover all alternatives,modifications, and equivalents, which may be included within the scopeof the present disclosure as defined by the embodiments.

Compounds

The “P” groups (e.g. P^(1a) and P^(1b)) defined for formula (I) hereinhave one bond to a —C(═O)— of formula (I) and one bond to a W^(1a)group. It is to be understood that a nitrogen of the P group isconnected to the —C(═O)— group of formula (I) and that a carbon of the Pgroup is connected to the W^(1a) group.

In the W^(1a) group a Y⁵ group is present. When that Y⁵ group is definedas —O—CH₂—, or —CH₂—O— group, those Y⁵ groups have a directionality. TheY⁵ group is connected to the W^(1a) group in the same left to rightdirectionality that each is drawn. So for example, when Y⁵ is —O—CH₂—,the directly following structure is intended:

For example, when Y⁵ is —CH₂—O—, the directly following structure isintended:

In the structure I, the W^(1a) group has a left-to-right directionalityas depicted in I and W^(1a) as they drawn.

E^(1a)-V^(1a)—C(═O)—P^(1a)—W^(1a)—P^(1b)—C(═O)—V^(1b)-E^(1b)  (I)

wherein:

W^(1a) is

For example, the P^(1a) group is connected to the imidazole group ofW^(1a), and the P1b group is connected to the pentacyclic ring system ofW^(1a).

“Alkyl” is C₁-C₁₈ hydrocarbon containing normal, secondary, tertiary orcyclic carbon atoms. Examples are methyl (Me, —CH₃), ethyl (Et,—CH₂CH₃), 1-propyl (n-Pr, n-propyl, —CH₂CH₂CH₃), 2-propyl (i-Pr,i-propyl, —CH(CH₃)₂), 1-butyl (n-Bu, n-butyl, —CH₂CH₂CH₂CH₃),2-methyl-1-propyl (i-Bu, i-butyl, —CH₂CH(CH₃)₂), 2-butyl (s-Bu, s-butyl,—CH(CH₃)CH₂CH₃), 2-methyl-2-propyl (t-Bu, t-butyl, —C(CH₃)₃), 1-pentyl(n-pentyl, —CH₂CH₂CH₂CH₂CH₃), 2-pentyl (—CH(CH₃)CH₂CH₂CH₃), 3-pentyl(—CH(CH₂CH₃)₂), 2-methyl-2-butyl (—C(CH₃)₂CH₂CH₃), 3-methyl-2-butyl(—CH(CH₃)CH(CH₃)₂), 3-methyl-1-butyl (—CH₂CH₂CH(CH₃)₂), 2-methyl-1-butyl(—CH₂CH(CH₃)CH₂CH₃), 1-hexyl (—CH₂CH₂CH₂CH₂CH₂CH₃), 2-hexyl(—CH(CH₃)CH₂CH₂CH₂CH₃), 3-hexyl (—CH(CH₂CH₃)(CH₂CH₂CH₃)),2-methyl-2-pentyl (—C(CH₃)₂CH₂CH₂CH₃), 3-methyl-2-pentyl(—CH(CH₃)CH(CH₃)CH₂CH₃), 4-methyl-2-pentyl (—CH(CH₃)CH₂CH(CH₃)₂),3-methyl-3-pentyl (—C(CH₃)(CH₂CH₃)₂), 2-methyl-3-pentyl(—CH(CH₂CH₃)CH(CH₃)₂), 2,3-dimethyl-2-butyl (—C(CH₃)₂CH(CH₃)₂),3,3-dimethyl-2-butyl (—CH(CH₃)C(CH₃)₃, and cyclopropylmethyl

“Alkenyl” is C₂-C₁₈ hydrocarbon containing normal, secondary, tertiaryor cyclic carbon atoms with at least one site of unsaturation, i.e. acarbon-carbon, sp² double bond. Examples include, but are not limitedto, ethylene or vinyl (—CH═CH₂), allyl (—CH₂CH═CH₂), cyclopentenyl(—C₅H₇), and 5-hexenyl (—CH₂CH₂CH₂CH₂CH═CH₂).

“Alkynyl” is C₂-C₁₈ hydrocarbon containing normal, secondary, tertiaryor cyclic carbon atoms with at least one site of unsaturation, i.e. acarbon-carbon, sp triple bond. Examples include, but are not limited to,acetylenic (—C≡CH) and propargyl (—CH₂C≡CH).

“Alkylene” refers to a saturated, branched or straight chain or cyclichydrocarbon radical of 1-18 carbon atoms, and having two monovalentradical centers derived by the removal of two hydrogen atoms from thesame or two different carbon atoms of a parent alkane. Typical alkyleneradicals include, but are not limited to, methylene (—CH₂—) 1,2-ethyl(—CH₂CH₂—), 1,3-propyl (—CH₂CH₂CH₂—), 1,4-butyl (—CH₂CH₂CH₂CH₂—), andthe like.

“Alkenylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical of 2-18 carbon atoms, and having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent alkene. Typicalalkenylene radicals include, but are not limited to, 1,2-ethylene(—CH═CH—).

“Alkynylene” refers to an unsaturated, branched or straight chain orcyclic hydrocarbon radical of 2-18 carbon atoms, and having twomonovalent radical centers derived by the removal of two hydrogen atomsfrom the same or two different carbon atoms of a parent alkyne. Typicalalkynylene radicals include, but are not limited to, acetylene (—C≡C—),propargyl (—CH₂C≡C—), and 4-pentynyl (—CH₂CH₂CH₂C≡CH).

The term “alkoxy” or “alkyloxy,” as used herein, refers to an alkylgroup attached to the parent molecular moiety through an oxygen atom.

The term “alkoxycarbonyl,” as used herein, refers to an alkoxy groupattached to the parent molecular moiety through a carbonyl group.

The term “cycloalkyl,” as used herein, refers to a saturated monocyclic,hydrocarbon ring system having three to seven carbon atoms and zeroheteroatoms. Representative examples of cycloalkyl groups include, butare not limited to, cyclopropyl, cyclopentyl, and cyclohexyl. Thecycloalkyl groups of the present disclosure are optionally substitutedwith one, two, three, four, or five substituents independently selectedfrom alkoxy, alkyl, aryl, cyano, halo, haloalkoxy, haloalkyl,heterocyclyl, hydroxy, hydroxyalkyl, nitro, and —NR^(x)R^(y) wherein thearyl and the heterocyclyl are further optionally substituted with one,two, or three substituents independently selected from alkoxy, alkyl,cyano, halo, haloalkoxy, haloalkyl, hydroxy, and nitro.

The term “cycloalkylcarbonyl,” as used herein, refers to a cycloalkylgroup attached to the parent molecular moiety through a carbonyl group.

The term “cycloalkyloxy,” as used herein, refers to a cycloalkyl groupattached to the parent molecular moiety through an oxygen atom.

The term “cycloalkyloxycarbonyl,” as used herein, refers to acycloalkyloxy group attached to the parent molecular moiety through acarbonyl group.

“Aryl” means a monovalent aromatic hydrocarbon radical of 6-20 carbonatoms derived by the removal of one hydrogen atom from a single carbonatom of a parent aromatic ring system. Typical aryl groups include, butare not limited to, radicals derived from benzene, substituted benzene,naphthalene, anthracene, biphenyl, and the like.

“Arylalkyl” refers to an acyclic alkyl radical in which one of thehydrogen atoms bonded to a carbon atom, typically a terminal or sp³carbon atom, is replaced with an aryl radical. Typical arylalkyl groupsinclude, but are not limited to, benzyl, 2-phenylethan-1-yl,naphthylmethyl, 2-naphthylethan-1-yl, naphthobenzyl,2-naphthophenylethan-1-yl and the like. The arylalkyl group comprises 6to 20 carbon atoms, e.g., the alkyl moiety, including alkanyl, alkenylor alkynyl groups, of the arylalkyl group is 1 to 6 carbon atoms and thearyl moiety is 5 to 14 carbon atoms.

“Substituted alkyl”, “substituted aryl”, and “substituted arylalkyl”mean alkyl, aryl, and arylalkyl respectively, in which one or morehydrogen atoms are each independently replaced with a non-hydrogensubstituent. Typical substituents include, but are not limited to: halo(e.g. F, Cl, Br, I), —R, —OR, —SR, —NR₂, —CF₃, —CCl₃, —OCF₃, —CN, —NO₂,—N(R)C(═O)R, —C(═O)R, —OC(═O)R, —C(O)OR, —C(═O)NRR, —S(═O)R, —S(═O)₂OR,—S(═O)₂R, —OS(═O)₂OR, —S(═O)₂NRR, and each R is independently —H, alkyl,aryl, arylalkyl, or heterocycle. Alkylene, alkenylene, and alkynylenegroups may also be similarly substituted.

The term “optionally substituted” in reference to a particular moiety ofthe compound of formula (I), (e.g., an optionally substituted arylgroup) refers to a moiety having 0, 1, 2, or more substituents.

The symbol “

” in a ring structure means that a bond is a single or double bond. In anon-limiting example,

can be

“Haloalkyl” as used herein includes an alkyl group substituted with oneor more halogens (e.g. F, Cl, Br, or I). Representative examples ofhaloalkyl include trifluoromethyl, 2,2,2-trifluoroethyl, and2,2,2-trifluoro-1-(trifluoromethyl)ethyl.

“Heterocycle” or “heterocyclyl” as used herein includes by way ofexample and not limitation these heterocycles described in Paquette, LeoA.; Principles of Modern Heterocyclic Chemistry (W.A. Benjamin, NewYork, 1968), particularly Chapters 1, 3, 4, 6, 7, and 9; The Chemistryof Heterocyclic Compounds, A Series of Monographs” (John Wiley & Sons,New York, 1950 to present), in particular Volumes 13, 14, 16, 19, and28; and J. Am. Chem. Soc. (1960) 82:5566. In one specific embodiment,“heterocycle” includes a “carbocycle” as defined herein, wherein one ormore (e.g. 1, 2, 3, or 4) carbon atoms have been replaced with aheteroatom (e.g. O, N, or S). The term heterocycle also includes“heteroaryl” which is a heterocycle wherein at least one heterocyclicrings is aromatic.

Examples of heterocycles include by way of example and not limitationpyridyl, dihydropyridyl, tetrahydropyridyl (piperidyl), thiazolyl,tetrahydrothiophenyl, sulfur oxidized tetrahydrothiophenyl, pyrimidinyl,furanyl, thienyl, pyrrolyl, pyrazolyl, imidazolyl, tetrazolyl,benzofuranyl, thianaphthalenyl, indolyl, indolenyl, quinolinyl,isoquinolinyl, benzimidazolyl, piperidinyl, 4-piperidonyl, pyrrolidinyl,2-pyrrolidonyl, pyrrolinyl, tetrahydrofuranyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, decahydroquinolinyl, octahydroisoquinolinyl,azocinyl, triazinyl, 6H-1,2,5-thiadiazinyl, 2H,6H-1,5,2-dithiazinyl,thienyl, thianthrenyl, pyranyl, isobenzofuranyl, chromenyl, xanthenyl,phenoxathinyl, 2H-pyrrolyl, isothiazolyl, isoxazolyl, pyrazinyl,pyridazinyl, indolizinyl, isoindolyl, 3H-indolyl, 1H-indazolyl, purinyl,4H-quinolizinyl, phthalazinyl, naphthyridinyl, quinoxalinyl,quinazolinyl, cinnolinyl, pteridinyl, 4H-carbazolyl, carbazolyl,β-carbolinyl, phenanthridinyl, acridinyl, pyrimidinyl, phenanthrolinyl,phenazinyl, phenothiazinyl, furazanyl, phenoxazinyl, isochromanyl,chromanyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl,piperazinyl, indolinyl, isoindolinyl, quinuclidinyl, morpholinyl,oxazolidinyl, benzotriazolyl, benzisoxazolyl, oxindolyl, benzoxazolinyl,isatinoyl, and bis-tetrahydrofuranyl:

By way of example and not limitation, carbon bonded heterocycles arebonded at position 2, 3, 4, 5, or 6 of a pyridine, position 3, 4, 5, or6 of a pyridazine, position 2, 4, 5, or 6 of a pyrimidine, position 2,3, 5, or 6 of a pyrazine, position 2, 3, 4, or 5 of a furan,tetrahydrofuran, thiofuran, thiophene, pyrrole or tetrahydropyrrole,position 2, 4, or 5 of an oxazole, imidazole or thiazole, position 3, 4,or 5 of an isoxazole, pyrazole, or isothiazole, position 2 or 3 of anaziridine, position 2, 3, or 4 of an azetidine, position 2, 3, 4, 5, 6,7, or 8 of a quinoline or position 1, 3, 4, 5, 6, 7, or 8 of anisoquinoline. Still more typically, carbon bonded heterocycles include2-pyridyl, 3-pyridyl, 4-pyridyl, 5-pyridyl, 6-pyridyl, 3-pyridazinyl,4-pyridazinyl, 5-pyridazinyl, 6-pyridazinyl, 2-pyrimidinyl,4-pyrimidinyl, 5-pyrimidinyl, 6-pyrimidinyl, 2-pyrazinyl, 3-pyrazinyl,5-pyrazinyl, 6-pyrazinyl, 2-thiazolyl, 4-thiazolyl, or 5-thiazolyl.

By way of example and not limitation, nitrogen bonded heterocycles arebonded at position 1 of an aziridine, azetidine, pyrrole, pyrrolidine,2-pyrroline, 3-pyrroline, imidazole, imidazolidine, 2-imidazoline,3-imidazoline, pyrazole, pyrazoline, 2-pyrazoline, 3-pyrazoline,piperidine, piperazine, indole, indoline, 1H-indazole, position 2 of aisoindole, or isoindoline, position 4 of a morpholine, and position 9 ofa carbazole, or β-carboline. Still more typically, nitrogen bondedheterocycles include 1-aziridyl, 1-azetedyl, 1-pyrrolyl, 1-imidazolyl,1-pyrazolyl, and 1-piperidinyl.

“Carbocycle” refers to a saturated, unsaturated or aromatic ring havingup to about 25 carbon atoms. Typically, a carbocycle has about 3 to 7carbon atoms as a monocycle, about 7 to 12 carbon atoms as a bicycle,and up to about 25 carbon atoms as a polycycle. Monocyclic carbocyclestypically have 3 to 6 ring atoms, still more typically 5 or 6 ringatoms. Bicyclic carbocycles typically have 7 to 12 ring atoms, e.g.,arranged as a bicyclo [4,5], [5,5], [5,6] or [6,6] system, or 9 or 10ring atoms arranged as a bicyclo [5,6] or [6,6] system. The termcarbocycle includes “cycloalkyl” which is a saturated or unsaturatedcarbocycle. Examples of monocyclic carbocycles include cyclopropyl,cyclobutyl, cyclopentyl, 1-cyclopent-1-enyl, 1-cyclopent-2-enyl,1-cyclopent-3-enyl, cyclohexyl, 1-cyclohex-1-enyl, 1-cyclohex-2-enyl,1-cyclohex-3-enyl, phenyl, spiryl and naphthyl.

The term “amino,” as used herein, refers to —NH₂.

The term “chiral” refers to molecules which have the property ofnon-superimposability of the mirror image partner, while the term“achiral” refers to molecules which are superimposable on their mirrorimage partner.

The term “stereoisomers” refers to compounds which have identicalchemical constitution, but differ with regard to the arrangement of theatoms or groups in space.

“Diastereomer” refers to a stereoisomer with two or more centers ofchirality and whose molecules are not mirror images of one another.Diastereomers have different physical properties, e.g., melting points,boiling points, spectral properties, and reactivities. Mixtures ofdiastereomers may separate under high resolution analytical proceduressuch as, for example, electrophoresis and chromatography.

“Enantiomers” refer to two stereoisomers of a compound which arenon-superimposable mirror images of one another.

The term “treatment” or “treating,” to the extent it relates to adisease or condition includes preventing the disease or condition fromoccurring, inhibiting the disease or condition, eliminating the diseaseor condition, and/or relieving one or more symptoms of the disease orcondition.

Stereochemical definitions and conventions used herein generally followS. P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984)McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S.,Stereochemistry of Organic Compounds (1994) John Wiley & Sons, Inc., NewYork. Many organic compounds exist in optically active forms, i.e., theyhave the ability to rotate the plane of plane-polarized light. Indescribing an optically active compound, the prefixes (D and L) or (Rand S) are used to denote the absolute configuration of the moleculeabout its chiral center(s). The prefixes d and 1 or (+) and (−) areemployed to designate the sign of rotation of plane-polarized light bythe compound, with (−) or 1 meaning that the compound is levorotatory. Acompound prefixed with (+) or d is dextrorotatory. For a given chemicalstructure, these stereoisomers are identical except that they are mirrorimages of one another. A specific stereoisomer may also be referred toas an enantiomer, and a mixture of such isomers is often called anenantiomeric mixture. A 50:50 mixture of enantiomers is referred to as aracemic mixture or a racemate, which may occur where there has been nostereoselection or stereospecificity in a chemical reaction or process.The terms “racemic mixture” and “racemate” refer to an equimolar mixtureof two enantiomeric species, devoid of optical activity. The disclosureincludes all stereoisomers of the compounds described herein.

Prodrugs

The term “prodrug” as used herein refers to any compound that whenadministered to a biological system generates a compound of thedisclosure that inhibits HCV activity (“the active inhibitorycompound”). The compound may be formed from the prodrug as a result of:(i) spontaneous chemical reaction(s), (ii) enzyme catalyzed chemicalreaction(s), (iii) photolysis, and/or (iv) metabolic chemicalreaction(s).

“Prodrug moiety” refers to a labile functional group which separatesfrom the active inhibitory compound during metabolism, systemically,inside a cell, by hydrolysis, enzymatic cleavage, or by some otherprocess (Bundgaard, Hans, “Design and Application of Prodrugs” in ATextbook of Drug Design and Development (1991), P. Krogsgaard-Larsen andH. Bundgaard, Eds. Harwood Academic Publishers, pp. 113-191). Enzymeswhich are capable of an enzymatic activation mechanism with the prodrugcompounds of the disclosure include, but are not limited to, amidases,esterases, microbial enzymes, phospholipases, cholinesterases, andphosphases. Prodrug moieties can serve to enhance solubility, absorptionand lipophilicity to optimize drug delivery, bioavailability andefficacy. A prodrug moiety may include an active metabolite or drugitself.

Exemplary prodrug moieties include the hydrolytically sensitive orlabile acyloxymethyl esters —CH₂OC(═O)R⁹⁹ and acyloxymethyl carbonates—CH₂OC(═O)OR⁹⁹ where R⁹⁹ is C₁-C₆ alkyl, C₁-C₆ substituted alkyl, C₆-C₂₀aryl or C₆-C₂₀ substituted aryl. The acyloxyalkyl ester was first usedas a prodrug strategy for carboxylic acids and then applied tophosphates and phosphonates by Farquhar et al. (1983) J. Pharm. Sci. 72:324; also U.S. Pat. Nos. 4,816,570, 4,968,788, 5,663,159 and 5,792,756.Subsequently, the acyloxyalkyl ester was used to deliver phosphonicacids across cell membranes and to enhance oral bioavailability. A closevariant of the acyloxyalkyl ester, the alkoxycarbonyloxyalkyl ester(carbonate), may also enhance oral bioavailability as a prodrug moietyin the compounds of the combinations of the disclosure. An exemplaryacyloxymethyl ester is pivaloyloxymethoxy, (POM) —CH₂OC(═O)C(CH₃)₃. Anexemplary acyloxymethyl carbonate prodrug moiety ispivaloyloxymethylcarbonate (POC) —CH₂OC(═O)OC(CH₃)₃.

Protecting Groups

In the context of the present disclosure, protecting groups includeprodrug moieties and chemical protecting groups.

“Protecting group” refers to a moiety of a compound that masks or altersthe properties of a functional group or the properties of the compoundas a whole. Chemical protecting groups and strategies forprotection/deprotection are well known in the art. See e.g., ProtectiveGroups in Organic Chemistry, Theodora W. Greene, John Wiley & Sons,Inc., New York, 1991. Protecting groups are often utilized to mask thereactivity of certain functional groups, to assist in the efficiency ofdesired chemical reactions, e.g., making and breaking chemical bonds inan ordered and planned fashion. Protection of functional groups of acompound alters other physical properties besides the reactivity of theprotected functional group, such as, for example, the polarity,lipophilicity (hydrophobicity), and other properties which can bemeasured by common analytical tools. Chemically protected intermediatesmay themselves be biologically active or inactive.

Protected compounds may also exhibit altered, and in some cases,optimized properties in vitro and in vivo, such as, for example, passagethrough cellular membranes and resistance to enzymatic degradation orsequestration. In this role, protected compounds with intendedtherapeutic effects may be referred to as prodrugs. Another function ofa protecting group is to convert the parental drug into a prodrug,whereby the parental drug is released upon conversion of the prodrug invivo. Because active prodrugs may be absorbed more effectively than theparental drug, prodrugs may possess greater potency in vivo than theparental drug. Protecting groups are removed either in vitro, in theinstance of chemical intermediates, or in vivo, in the case of prodrugs.With chemical intermediates, it is not particularly important that theresulting products after deprotection, e.g., alcohols, bephysiologically acceptable, although in general it is more desirable ifthe products are pharmacologically innocuous.

Protecting groups are available, commonly known and used, and areoptionally used to prevent side reactions with the protected groupduring synthetic procedures, i.e. routes or methods to prepare thecompounds of the disclosure. For the most part the decision as to whichgroups to protect, when to do so, and the nature of the chemicalprotecting group “PG” will be dependent upon the chemistry of thereaction to be protected against (e.g., acidic, basic, oxidative,reductive or other conditions) and the intended direction of thesynthesis. PGs do not need to be, and generally are not, the same if thecompound is substituted with multiple PG. In general, PG will be used toprotect functional groups such as, for example, carboxyl, hydroxyl,thio, or amino groups and to thus prevent side reactions or to otherwisefacilitate the synthetic efficiency. The order of deprotection to yieldfree deprotected groups is dependent upon the intended direction of thesynthesis and the reaction conditions to be encountered, and may occurin any order as determined by the artisan.

Various functional groups of the compounds of the disclosure may beprotected. For example, protecting groups for —OH groups (whetherhydroxyl, carboxylic acid, phosphonic acid, or other functions) include“ether- or ester-forming groups”. Ether- or ester-forming groups arecapable of functioning as chemical protecting groups in the syntheticschemes set forth herein. However, some hydroxyl and thio protectinggroups are neither ether—nor ester-forming groups, as will be understoodby those skilled in the art, and are included with amides, discussedbelow.

A very large number of hydroxyl protecting groups and amide-forminggroups and corresponding chemical cleavage reactions are described inProtective Groups in Organic Synthesis, Theodora W. Greene (John Wiley &Sons, Inc., New York, 1991, ISBN 0-471-62301-6) (“Greene”). See alsoKocienski, Philip J.; Protecting Groups (Georg Thieme Verlag Stuttgart,New York, 1994), which is incorporated by reference in its entiretyherein. In particular Chapter 1, Protecting Groups: An Overview, pages1-20, Chapter 2, Hydroxyl Protecting Groups, pages 21-94, Chapter 3,Diol Protecting Groups, pages 95-117, Chapter 4, Carboxyl ProtectingGroups, pages 118-154, Chapter 5, Carbonyl Protecting Groups, pages155-184. For protecting groups for carboxylic acid, phosphonic acid,phosphonate, sulfonic acid and other protecting groups for acids seeGreene as set forth below.

Stereoisomers

The compounds of the disclosure may have chiral centers, e.g., chiralcarbon or phosphorus atoms. The compounds of the disclosure thus includeall stereoisomers, including enantiomers, diastereomers, andatropisomers. In addition, the compounds of the disclosure includeenriched or resolved optical isomers at any or all asymmetric, chiralatoms. In other words, the chiral centers apparent from the depictionsare provided as the non-racemic or racemic mixtures. Both racemic anddiastereomeric mixtures, as well as the individual optical isomersisolated or synthesized, substantially free of their enantiomeric ordiastereomeric partners, are all within the scope of the disclosure. Theracemic mixtures are separated into their individual, substantiallyoptically pure isomers through well-known techniques such as, forexample, the separation of diastereomeric salts formed with opticallyactive adjuncts, e.g., acids or bases followed by conversion back to theoptically active substances. In most instances, the desired opticalisomer is synthesized by means of stereospecific reactions, beginningwith the appropriate stereoisomer of the desired starting material orthrough enantioselective reactions.

The compounds of the disclosure can also exist as tautomeric isomers incertain cases. Although only one tautomer may be depicted, all suchforms are contemplated within the scope of the disclosure. For example,ene-amine tautomers can exist for purine, pyrimidine, imidazole,guanidine, amidine, and tetrazole systems and all their possibletautomeric forms are within the scope of the disclosure.

Salts and Hydrates

Examples of physiologically or pharmaceutically acceptable salts of thecompounds of the disclosure include salts derived from an appropriatebase, such as, for example, an alkali metal (for example, sodium), analkaline earth metal (for example, magnesium), ammonium and NX₄ ⁺(wherein X is C₁-C₄ alkyl). Physiologically acceptable salts of ahydrogen atom or an amino group include salts of organic carboxylicacids such as, for example, acetic, benzoic, lactic, fumaric, tartaric,maleic, malonic, malic, isethionic, lactobionic and succinic acids;organic sulfonic acids, such as, for example, methanesulfonic,ethanesulfonic, benzenesulfonic and p-toluenesulfonic acids; andinorganic acids, such as, for example, hydrochloric, sulfuric,phosphoric and sulfamic acids. Physiologically acceptable salts of acompound of a hydroxy group include the anion of said compound incombination with a suitable cation such as, for example, Na⁺ and NX₄ ⁺(wherein X is independently selected from H or a C₁-C₄ alkyl group).

For therapeutic use, salts of active ingredients of the compounds of thedisclosure will typically be physiologically acceptable, i.e. they willbe salts derived from a physiologically acceptable acid or base.However, salts of acids or bases which are not physiologicallyacceptable may also find use, for example, in the preparation orpurification of a physiologically acceptable compound. All salts,whether or not derived form a physiologically acceptable acid or base,are within the scope of the present disclosure.

Metal salts typically are prepared by reacting the metal hydroxide witha compound of this disclosure. Examples of metal salts which areprepared in this way are salts containing Li⁺, Na⁺, and K⁺. A lesssoluble metal salt can be precipitated from the solution of a moresoluble salt by addition of the suitable metal compound.

In addition, salts may be formed from acid addition of certain organicand inorganic acids, e.g., HCl, HBr, H₂SO₄, H₃PO₄ or organic sulfonicacids, to basic centers, typically amines, or to acidic groups. Finally,it is to be understood that the compositions herein comprise compoundsof the disclosure in their un-ionized, as well as zwitterionic form, andcombinations with stoichiometric amounts of water as in hydrates.

Also included within the scope of this disclosure are the salts of theparental compounds with one or more amino acids. Any of the natural orunnatural amino acids are suitable, especially the naturally-occurringamino acids found as protein components, although the amino acidtypically is one bearing a side chain with a basic or acidic group,e.g., lysine, arginine or glutamic acid, or a neutral group such as, forexample, glycine, serine, threonine, alanine, isoleucine, or leucine.

Methods of Inhibition of HCV

Another aspect of the disclosure relates to methods of inhibiting theactivity of HCV comprising the step of treating a sample suspected ofcontaining HCV with a compound or composition of the disclosure.

The treating step of the disclosure comprises adding the compound of thedisclosure to the sample or it comprises adding a precursor of thecomposition to the sample. The addition step comprises any method ofadministration as described above.

If desired, the activity of HCV after application of the compound can beobserved by any method including direct and indirect methods ofdetecting HCV activity. Quantitative, qualitative, and semiquantitativemethods of determining HCV activity are all contemplated. Typically oneof the screening methods described above are applied, however, any othermethod such as, for example, observation of the physiological propertiesof a living organism are also applicable.

Many organisms contain HCV. The compounds of this disclosure are usefulin the treatment or prophylaxis of conditions associated with HCVactivation in animals or in man.

However, in screening compounds capable of inhibiting HCV activity itshould be kept in mind that the results of enzyme assays may not alwayscorrelate with cell culture assays. Thus, a cell based assay shouldtypically be the primary screening tool.

Pharmaceutical Formulations

The compounds of this disclosure are formulated with conventionalcarriers and excipients, which will be selected in accord with ordinarypractice. Tablets will contain excipients, glidants, fillers, bindersand the like. Aqueous formulations are prepared in sterile form, andwhen intended for delivery by other than oral administration generallywill be isotonic. All formulations will optionally contain excipientssuch as, for example, those set forth in the Handbook of PharmaceuticalExcipients (1986). Excipients include ascorbic acid and otherantioxidants, chelating agents such as, for example, EDTA, carbohydratessuch as, for example, dextrin, hydroxyalkylcellulose,hydroxyalkylmethylcellulose, stearic acid and the like. The pH of theformulations ranges from about 3 to about 11, but is ordinarily about 7to 10. Typically, the compound will be administered in a dose from 0.01milligrams to 2 grams. In one embodiment, the dose will be from about 10milligrams to 450 milligrams. It is contemplated that the compound maybe administered once, twice or three times a day.

While it is possible for the active ingredients to be administered aloneit may be preferable to present them as pharmaceutical formulations. Theformulations, both for veterinary and for human use, of the disclosurecomprise at least one active ingredient, as above defined, together withone or more acceptable carriers therefore and optionally othertherapeutic ingredients. The carrier(s) must be “acceptable” in thesense of being compatible with the other ingredients of the formulationand physiologically innocuous to the recipient thereof.

The formulations include those suitable for the foregoing administrationroutes. The formulations may conveniently be presented in unit dosageform and may be prepared by any of the methods well known in the art ofpharmacy. Techniques and formulations generally are found in Remington'sPharmaceutical Sciences (Mack Publishing Co., Easton, Pa.). Such methodsinclude the step of bringing into association the active ingredient withthe carrier which constitutes one or more accessory ingredients. Ingeneral the formulations are prepared by uniformly and intimatelybringing into association the active ingredient with liquid carriers orfinely divided solid carriers or both, and then, if necessary, shapingthe product.

Formulations of the present disclosure suitable for oral administrationmay be presented as discrete units such as, for example, capsules,cachets or tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion ora water-in-oil liquid emulsion. The active ingredient may also beadministered as a bolus, electuary or paste.

A tablet is made by compression or molding, optionally with one or moreaccessory ingredients. Compressed tablets may be prepared by compressingin a suitable machine the active ingredient in a free-flowing form suchas, for example, a powder or granules, optionally mixed with a binder,lubricant, inert diluent, preservative, surface active or dispersingagent. Molded tablets may be made by molding in a suitable machine amixture of the powdered active ingredient moistened with an inert liquiddiluent. The tablets may optionally be coated or scored and optionallyare formulated so as to provide slow or controlled release of the activeingredient therefrom.

For administration to the eye or other external tissues e.g., mouth andskin, the formulations are preferably applied as a topical ointment orcream containing the active ingredient(s) in an amount of, for example,0.075 to 20% w/w (including active ingredient(s) in a range between 0.1%and 20% in increments of 0.1% w/w such as, for example, 0.6% w/w, 0.7%w/w, etc.), preferably 0.2 to 15% w/w and most preferably 0.5 to 10%w/w. When formulated in an ointment, the active ingredients may beemployed with either a paraffinic or a water-miscible ointment base.Alternatively, the active ingredients may be formulated in a cream withan oil-in-water cream base.

If desired, the aqueous phase of the cream base may include, forexample, at least 30% w/w of a polyhydric alcohol, i.e. an alcoholhaving two or more hydroxyl groups such as, for example, propyleneglycol, butane 1,3-diol, mannitol, sorbitol, glycerol and polyethyleneglycol (including PEG 400) and mixtures thereof. The topicalformulations may desirably include a compound which enhances absorptionor penetration of the active ingredient through the skin or otheraffected areas. Examples of such dermal penetration enhancers includedimethyl sulphoxide and related analogs.

The oily phase of the emulsions of this disclosure may be constitutedfrom known ingredients in a known manner. While the phase may comprisemerely an emulsifier (otherwise known as an emulgent), it desirablycomprises a mixture of at least one emulsifier with a fat or an oil orwith both a fat and an oil. Preferably, a hydrophilic emulsifier isincluded together with a lipophilic emulsifier which acts as astabilizer. It is also preferred to include both an oil and a fat.Together, the emulsifier(s) with or without stabilizer(s) make up theso-called emulsifying wax, and the wax together with the oil and fatmake up the so-called emulsifying ointment base which forms the oilydispersed phase of the cream formulations.

Emulgents and emulsion stabilizers suitable for use in the formulationof the disclosure include Tween® 60, Span® 80, cetostearyl alcohol,benzyl alcohol, myristyl alcohol, glyceryl mono-stearate and sodiumlauryl sulfate.

The choice of suitable oils or fats for the formulation is based onachieving the desired cosmetic properties. The cream should preferablybe a non-greasy, non-staining and washable product with suitableconsistency to avoid leakage from tubes or other containers. Straight orbranched chain, mono- or dibasic alkyl esters such as, for example,di-isoadipate, isocetyl stearate, propylene glycol diester of coconutfatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate,butyl stearate, 2-ethylhexyl palmitate or a blend of branched chainesters known as Crodamol CAP may be used, the last three being preferredesters. These may be used alone or in combination depending on theproperties required. Alternatively, high melting point lipids such as,for example, white soft paraffin and/or liquid paraffin or other mineraloils are used.

Pharmaceutical formulations according to the present disclosure compriseone or more compounds of the disclosure together with one or morepharmaceutically acceptable carriers or excipients and optionally othertherapeutic agents. Pharmaceutical formulations containing the activeingredient may be in any form suitable for the intended method ofadministration. When used for oral use for example, tablets, troches,lozenges, aqueous or oil suspensions, dispersible powders or granules,emulsions, hard or soft capsules, syrups or elixirs may be prepared.Compositions intended for oral use may be prepared according to anymethod known to the art for the manufacture of pharmaceuticalcompositions and such compositions may contain one or more agentsincluding sweetening agents, flavoring agents, coloring agents andpreserving agents, in order to provide a palatable preparation. Tabletscontaining the active ingredient in admixture with non-toxicpharmaceutically acceptable excipient which are suitable for manufactureof tablets are acceptable. These excipients may be, for example, inertdiluents, such as, for example, calcium or sodium carbonate, lactose,lactose monohydrate, croscarmellose sodium, povidone, calcium or sodiumphosphate; granulating and disintegrating agents, such as, for example,maize starch, or alginic acid; binding agents, such as, for example,cellulose, microcrystalline cellulose, starch, gelatin or acacia; andlubricating agents, such as, for example, magnesium stearate, stearicacid or talc. Tablets may be uncoated or may be coated by knowntechniques including microencapsulation to delay disintegration andadsorption in the gastrointestinal tract and thereby provide a sustainedaction over a longer period. For example, a time delay material such as,for example, glyceryl monostearate or glyceryl distearate alone or witha wax may be employed.

Formulations for oral use may be also presented as hard gelatin capsuleswhere the active ingredient is mixed with an inert solid diluent, forexample calcium phosphate or kaolin, or as soft gelatin capsules whereinthe active ingredient is mixed with water or an oil medium, such as, forexample, peanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the disclosure contain the active materials inadmixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include a suspending agent, such as, forexample, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcelluose, sodium alginate, polyvinylpyrrolidone, gum tragacanthand gum acacia, and dispersing or wetting agents such as, for example, anaturally occurring phosphatide (e.g., lecithin), a condensation productof an alkylene oxide with a fatty acid (e.g., polyoxyethylene stearate),a condensation product of ethylene oxide with a long chain aliphaticalcohol (e.g., heptadecaethyleneoxycetanol), a condensation product ofethylene oxide with a partial ester derived from a fatty acid and ahexitol anhydride (e.g., polyoxyethylene sorbitan monooleate). Theaqueous suspension may also contain one or more preservatives such as,for example, ethyl or n-propyl p-hydroxy-benzoate, one or more coloringagents, one or more flavoring agents and one or more sweetening agents,such as, for example, sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient ina vegetable oil, such as, for example, arachis oil, olive oil, sesameoil or coconut oil, or in a mineral oil such as, for example, liquidparaffin. The oral suspensions may contain a thickening agent, such as,for example, beeswax, hard paraffin or cetyl alcohol. Sweetening agents,such as, for example, those set forth above, and flavoring agents may beadded to provide a palatable oral preparation. These compositions may bepreserved by the addition of an antioxidant such as, for example,ascorbic acid.

Dispersible powders and granules of the disclosure suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Suitable dispersing orwetting agents and suspending agents are exemplified by those disclosedabove. Additional excipients, for example sweetening, flavoring andcoloring agents, may also be present.

The pharmaceutical compositions of the disclosure may also be in theform of oil-in-water emulsions. The oily phase may be a vegetable oil,such as, for example, olive oil or arachis oil, a mineral oil, such as,for example, liquid paraffin, or a mixture of these. Suitableemulsifying agents include naturally-occurring gums, such as, forexample, gum acacia and gum tragacanth, naturally occurringphosphatides, such as, for example, soybean lecithin, esters or partialesters derived from fatty acids and hexitol anhydrides, such as, forexample, sorbitan monooleate, and condensation products of these partialesters with ethylene oxide, such as, for example, polyoxyethylenesorbitan monooleate. The emulsion may also contain sweetening andflavoring agents. Syrups and elixirs may be formulated with sweeteningagents, such as, for example, glycerol, sorbitol or sucrose. Suchformulations may also contain a demulcent, a preservative, a flavoringor a coloring agent.

The pharmaceutical compositions of the disclosure may be in the form ofa sterile injectable preparation, such as, for example, a sterileinjectable aqueous or oleaginous suspension. This suspension may beformulated according to the known art using those suitable dispersing orwetting agents and suspending agents which have been mentioned above.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, such as, for example, a solution in 1,3-butane-diol or preparedas a lyophilized powder. Among the acceptable vehicles and solvents thatmay be employed are water, Ringer's solution and isotonic sodiumchloride solution. In addition, sterile fixed oils may conventionally beemployed as a solvent or suspending medium. For this purpose any blandfixed oil may be employed including synthetic mono- or diglycerides. Inaddition, fatty acids such as, for example, oleic acid may likewise beused in the preparation of injectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain approximately 1 to 1000 mg of active material compounded with anappropriate and convenient amount of carrier material which may varyfrom about 5 to about 95% of the total compositions (weight:weight). Thepharmaceutical composition can be prepared to provide easily measurableamounts for administration. For example, an aqueous solution intendedfor intravenous infusion may contain from about 3 to 500 μg of theactive ingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/hr can occur.

Formulations suitable for administration to the eye include eye dropswherein the active ingredient is dissolved or suspended in a suitablecarrier, especially an aqueous solvent for the active ingredient. Theactive ingredient is preferably present in such formulations in aconcentration of 0.5 to 20%, advantageously 0.5 to 10% particularlyabout 1.5% w/w.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored basis, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert basis such as, for example, gelatin and glycerin,or sucrose and acacia; and mouthwashes comprising the active ingredientin a suitable liquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for intrapulmonary or nasal administration have aparticle size for example in the range of 0.1 to 500 microns (includingparticle sizes in a range between 0.1 and 500 microns in incrementsmicrons such as, for example, 0.5, 1, 30 microns, 35 microns, etc.),which is administered by rapid inhalation through the nasal passage orby inhalation through the mouth so as to reach the alveolar sacs.Suitable formulations include aqueous or oily solutions of the activeingredient. Formulations suitable for aerosol or dry powderadministration may be prepared according to conventional methods and maybe delivered with other therapeutic agents such as, for example,compounds heretofore used in the treatment or prophylaxis of conditionsassociated with HCV activity.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

Formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats and solutes which render the formulation isotonicwith the blood of the intended recipient; and aqueous and non-aqueoussterile suspensions which may include suspending agents and thickeningagents.

The formulations are presented in unit-dose or multi-dose containers,for example sealed ampoules and vials, and may be stored in afreeze-dried (lyophilized) condition requiring only the addition of thesterile liquid carrier, for example water for injection, immediatelyprior to use. Extemporaneous injection solutions and suspensions areprepared from sterile powders, granules and tablets of the kindpreviously described. Preferred unit dosage formulations are thosecontaining a daily dose or unit daily sub-dose, as herein above recited,or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the ingredients particularlymentioned above the formulations of this disclosure may include otheragents conventional in the art having regard to the type of formulationin question, for example those suitable for oral administration mayinclude flavoring agents.

The disclosure further provides veterinary compositions comprising atleast one active ingredient as above defined together with a veterinarycarrier therefore.

Veterinary carriers are materials useful for the purpose ofadministering the composition and may be solid, liquid or gaseousmaterials which are otherwise inert or acceptable in the veterinary artand are compatible with the active ingredient. These veterinarycompositions may be administered orally, parenterally or by any otherdesired route.

Compounds of the disclosure can also be formulated to provide controlledrelease of the active ingredient to allow less frequent dosing or toimprove the pharmacokinetic or toxicity profile of the activeingredient. Accordingly, the disclosure also provides compositionscomprising one or more compounds of the disclosure formulated forsustained or controlled release.

Effective dose of active ingredient depends at least on the nature ofthe condition being treated, toxicity, whether the compound is beingused prophylactically (lower doses), the method of delivery, and thepharmaceutical formulation, and will be determined by the clinicianusing conventional dose escalation studies.

Routes of Administration

One or more compounds of the disclosure (herein referred to as theactive ingredients) are administered by any route appropriate to thecondition to be treated. Suitable routes include oral, rectal, nasal,topical (including buccal and sublingual), vaginal and parenteral(including subcutaneous, intramuscular, intravenous, intradermal,intrathecal and epidural), and the like. It will be appreciated that thepreferred route may vary with for example the condition of therecipient. An advantage of the compounds of this disclosure is that theyare orally bioavailable and can be dosed orally.

HCV Combination Therapy

In another embodiment, non-limiting examples of suitable combinationsinclude combinations of one or more compounds of formula (I) and (A1-A4)with one or more interferons, ribavirin or its analogs, HCV NS3 proteaseinhibitors, alpha-glucosidase 1 inhibitors, hepatoprotectants,nucleoside or nucleotide inhibitors of HCV NS5B polymerase,non-nucleoside inhibitors of HCV NS5B polymerase, HCV NS5A inhibitors,TLR-7 agonists, cyclophillin inhibitors, HCV IRES inhibitors,pharmacokinetic enhancers, and other drugs or therapeutic agents fortreating HCV.

More specifically, one or more compounds of the present as describedherein may be combined with one or more compounds selected from thegroup consisting of

1) interferons, e.g., pegylated rIFN-alpha 2b (PEG-Intron®), pegylatedrIFN-alpha 2a (Pegasys®), rIFN-alpha 2b (Intron® A), rIFN-alpha 2a(Roferon®-A), interferon alpha (MOR-22, OPC-18, Alfaferone®,Alfanative®, Multiferon®, subalin), interferon alfacon-1 (Infergen®),interferon alpha-n1 (Wellferon), interferon alpha-n3 (Alferon®),interferon-beta (Avonex®, DL-8234), interferon-omega (omega DUROS®,Biomed® 510), albinterferon alpha-2b (Albuferon®), IFN alpha-2b XL,BLX-883 (Locteron®), DA-3021, glycosylated interferon alpha-2b(AVI-005), PEG-Infergen, PEGylated interferon lambda-1 (PEGylatedIL-29), and Belerofon®;

2) ribavirin and its analogs, e.g., ribavirin (Rebetol®, Copegus®), andtaribavirin (Viramidine®);

3) HCV NS3 protease inhibitors, e.g., boceprevir (SCH-503034, SCH-7),telaprevir (VX-950), TMC435350, BI-1335, BI-1230, MK-7009, VBY-376,VX-500, GS-9256, GS-9451, BMS-605339, PHX-1766, AS-101, YH-5258, YH5530,YH5531, ABT-450, ACH-1625, ITMN-191, AT26893, MK5172, MK6325, andMK2748;

4) alpha-glucosidase 1 inhibitors, e.g., celgosivir (MX-3253), Miglitol,and UT-231B;

5) hepatoprotectants, e.g., emericasan (IDN-6556), ME-3738, GS-9450(LB-84451), silibilin, and MitoQ;

6) nucleoside or nucleotide inhibitors of HCV NS5B polymerase, e.g.,R1626, R7128 (R4048), IDX184, IDX-102, BCX-4678, valopicitabine(NM-283), MK-0608, sofosbuvir (GS-7977 (formerly PSI-7977)), VLX-135(formerly ALS-2200), and INX-189 (now BMS986094);

7) non-nucleoside inhibitors of HCV NS5B polymerase, e.g., PF-868554,VCH-759, VCH-916, JTK-652, MK-3281, GS-9190, VBY-708, VCH-222, A848837,ANA-598, GL60667, GL59728, A-63890, A-48773, A-48547, BC-2329, VCH-796(nesbuvir), GSK625433, BILN-1941, XTL-2125, ABT-072, ABT-333, GS-9669,PSI-7792, and GS-9190;

8) HCV NS5A inhibitors, e.g., AZD-2836 (A-831), BMS-790052, ACH-3102,ACH-2928, MK8325, MK4882, MK8742, PSI-461, IDX719, GS-5885, and A-689;

9) TLR-7 agonists, e.g., imiquimod, 852A, GS-9524, ANA-773, ANA-975(isatoribine), AZD-8848 (DSP-3025), and SM-360320;

10) cyclophillin inhibitors, e.g., DEBIO-025, SCY-635, and NIM811;

11) HCV IRES inhibitors, e.g., MCI-067;

12) pharmacokinetic enhancers, e.g., BAS-100, SPI-452, PF-4194477,TMC-41629, GS-9350 (cobicistat), GS-9585, and roxythromycin; and

13) other drugs for treating HCV, e.g., thymosin alpha 1 (Zadaxin),nitazoxanide (Alinea, NTZ), BIVN-401 (virostat), PYN-17 (altirex),KPE02003002, actilon (CPG-10101), GS-9525, KRN-7000, civacir, GI-5005,XTL-6865, BIT225, PTX-111, ITX2865, TT-033i, ANA 971, NOV-205, tarvacin,EHC-18, VGX-410C, EMZ-702, AVI 4065, BMS-650032, BMS-791325,Bavituximab, MDX-1106 (ONO-4538), Oglufanide, and VX-497 (merimepodib).

In yet another embodiment, the present application disclosespharmaceutical compositions comprising a compound as described herein,or a pharmaceutically acceptable salt, solvate, and/or ester thereof, incombination with at least one additional therapeutic agent, and apharmaceutically acceptable carrier or excipient.

In another embodiment is provided a pharmaceutical compositioncomprising a compound of formula (I) as described herein and sofosbuvirand/or GS-5885 and optionally an interferon or ribavirin.

It is contemplated that additional therapeutic agents will beadministered in a manner that is known in the art and the dosage may beselected by someone of skill in the art. For example, additionaltherapeutic agents may be administered in a dose from about 0.01milligrams to about 2 grams per day.

Metabolites of the Compounds

Also falling within the scope of this disclosure are the in vivometabolic products of the compounds described herein. Such products mayresult for example from the oxidation, reduction, hydrolysis, amidation,esterification and the like of the administered compound, primarily dueto enzymatic processes. Accordingly, the disclosure includes compoundsproduced by a process comprising contacting a compound of thisdisclosure with a mammal for a period of time sufficient to yield ametabolic product thereof. Such products typically are identified bypreparing a radiolabelled (e.g., C¹⁴ or H³) compound of the disclosure,administering it parenterally in a detectable dose (e.g., greater thanabout 0.5 mg/kg) to an animal such as, for example, rat, mouse, guineapig, monkey, or to man, allowing sufficient time for metabolism to occur(typically about 30 seconds to 30 hours) and isolating its conversionproducts from the urine, blood or other biological samples. Theseproducts are easily isolated since they are labeled (others are isolatedby the use of antibodies capable of binding epitopes surviving in themetabolite). The metabolite structures are determined in conventionalfashion, e.g., by MS or NMR analysis. In general, analysis ofmetabolites is done in the same way as conventional drug metabolismstudies well-known to those skilled in the art. The conversion products,so long as they are not otherwise found in vivo, are useful indiagnostic assays for therapeutic dosing of the compounds of thedisclosure even if they possess no HCV-inhibitory activity of their own.

Methods for determining stability of compounds in surrogategastrointestinal secretions are known.

Exemplary Methods of Making the Compounds

The disclosure also relates to methods of making the compositions of thedisclosure. The compositions are prepared by any of the applicabletechniques of organic synthesis. Many such techniques are well known inthe art. However, many of the known techniques are elaborated inCompendium of Organic Synthetic Methods (John Wiley & Sons, New York),Vol. 1, Ian T. Harrison and Shuyen Harrison, 1971; Vol. 2, Ian T.Harrison and Shuyen Harrison, 1974; Vol. 3, Louis S. Hegedus and LeroyWade, 1977; Vol. 4, Leroy G. Wade, Jr., 1980; Vol. 5, Leroy G. Wade,Jr., 1984; and Vol. 6, Michael B. Smith; as well as March, J., AdvancedOrganic Chemistry, Third Edition, (John Wiley & Sons, New York, 1985),Comprehensive Organic Synthesis. Selectivity, Strategy & Efficiency inModern Organic Chemistry. In 9 Volumes, Barry M. Trost, Editor-in-Chief(Pergamon Press, New York, 1993 printing). Other methods suitable forpreparing compounds of the disclosure are described in InternationalPatent Application Publication Number WO 2006/020276.

A number of exemplary methods for the preparation of the compositions ofthe disclosure are provided in the schemes and examples below. Thesemethods are intended to illustrate the nature of such preparations andare not intended to limit the scope of applicable methods.

Generally, the reaction conditions such as, for example, temperature,reaction time, solvents, work-up procedures, and the like, will be thosecommon in the art for the particular reaction to be performed. The citedreference material, together with material cited therein, containsdetailed descriptions of such conditions. Typically the temperatureswill be −100° C. to 200° C., solvents will be aprotic or protic, andreaction times will be 10 seconds to 10 days. Work-up typically consistsof quenching any unreacted reagents followed by partition between awater/organic layer system (extraction) and separating the layercontaining the product.

Oxidation and reduction reactions are typically carried out attemperatures near room temperature (about 20° C.), although for metalhydride reductions frequently the temperature is reduced to 0° C. to−100° C., solvents are typically aprotic for reductions and may beeither protic or aprotic for oxidations. Reaction times are adjusted toachieve desired conversions.

Condensation reactions are typically carried out at temperatures nearroom temperature, although for non-equilibrating, kinetically controlledcondensations reduced temperatures (0° C. to −100° C.) are also common.Solvents can be either protic (common in equilibrating reactions) oraprotic (common in kinetically controlled reactions).

Standard synthetic techniques such as, for example, azeotropic removalof reaction by-products and use of anhydrous reaction conditions (e.g.,inert gas environments) are common in the art and will be applied whenapplicable.

The terms “treated”, “treating”, “treatment”, and the like, when used inconnection with a chemical synthetic operation, mean contacting, mixing,reacting, allowing to react, bringing into contact, and other termscommon in the art for indicating that one or more chemical entities istreated in such a manner as to convert it to one or more other chemicalentities. This means that “treating compound one with compound two” issynonymous with “allowing compound one to react with compound two”,“contacting compound one with compound two”, “reacting compound one withcompound two”, and other expressions common in the art of organicsynthesis for reasonably indicating that compound one was “treated”,“reacted”, “allowed to react”, etc., with compound two. For example,treating indicates the reasonable and usual manner in which organicchemicals are allowed to react. Normal concentrations (0.01M to 10M,typically 0.1M to 1M), temperatures (−100° C. to 250° C., typically −78°C. to 150° C., more typically −78° C. to 100° C., still more typically0° C. to 100° C.), reaction vessels (typically glass, plastic, metal),solvents, pressures, atmospheres (typically air for oxygen and waterinsensitive reactions or nitrogen or argon for oxygen or watersensitive), etc., are intended unless otherwise indicated. The knowledgeof similar reactions known in the art of organic synthesis is used inselecting the conditions and apparatus for “treating” in a givenprocess. In particular, one of ordinary skill in the art of organicsynthesis selects conditions and apparatus reasonably expected tosuccessfully carry out the chemical reactions of the described processesbased on the knowledge in the art.

Modifications of each of the exemplary schemes and in the Examples(hereafter “exemplary schemes”) leads to various analogs of the specificexemplary materials produce. The above-cited citations describingsuitable methods of organic synthesis are applicable to suchmodifications.

In each of the exemplary schemes it may be advantageous to separatereaction products from one another and/or from starting materials. Thedesired products of each step or series of steps is separated and/orpurified (hereinafter separated) to the desired degree of homogeneity bythe techniques common in the art. Typically such separations involvemultiphase extraction, crystallization from a solvent or solventmixture, distillation, sublimation, or chromatography. Chromatographycan involve any number of methods including, for example: reverse-phaseand normal phase; size exclusion; ion exchange; high, medium, and lowpressure liquid chromatography methods and apparatus; small scaleanalytical; simulated moving bed (SMB) and preparative thin or thicklayer chromatography, as well as techniques of small scale thin layerand flash chromatography.

Another class of separation methods involves treatment of a mixture witha reagent selected to bind to or render otherwise separable a desiredproduct, unreacted starting material, reaction by product, or the like.Such reagents include adsorbents or absorbents such as, for example,activated carbon, molecular sieves, ion exchange media, or the like.Alternatively, the reagents can be acids in the case of a basicmaterial, bases in the case of an acidic material, binding reagents suchas, for example, antibodies, binding proteins, selective chelators suchas, for example, crown ethers, liquid/liquid ion extraction reagents(LIX), or the like.

Selection of appropriate methods of separation depends on the nature ofthe materials involved. For example, boiling point, and molecular weightin distillation and sublimation, presence or absence of polar functionalgroups in chromatography, stability of materials in acidic and basicmedia in multiphase extraction, and the like. One skilled in the artwill apply techniques most likely to achieve the desired separation.

A single stereoisomer, e.g., an enantiomer, substantially free of itsstereoisomer may be obtained by resolution of the racemic mixture usinga method such as, for example, formation of diastereomers usingoptically active resolving agents (Stereochemistry of Carbon Compounds,(1962) by E. L. Eliel, McGraw Hill; Lochmuller, C. H., (1975) J.Chromatogr., 113, 3) 283-302). Racemic mixtures of chiral compounds ofthe disclosure can be separated and isolated by any suitable method,including: (1) formation of ionic, diastereomeric salts with chiralcompounds and separation by fractional crystallization or other methods,(2) formation of diastereomeric compounds with chiral derivatizingreagents, separation of the diastereomers, and conversion to the purestereoisomers, and (3) separation of the substantially pure or enrichedstereoisomers directly under chiral conditions.

Under method (1), diastereomeric salts can be formed by reaction ofenantiomerically pure chiral bases such as, for example, brucine,quinine, ephedrine, strychnine, α-methyl-β-phenylethylamine(amphetamine), and the like with asymmetric compounds bearing acidicfunctionality, such as, for example, carboxylic acid and sulfonic acid.The diastereomeric salts may be induced to separate by fractionalcrystallization or ionic chromatography. For separation of the opticalisomers of amino compounds, addition of chiral carboxylic or sulfonicacids, such as, for example, camphorsulfonic acid, tartaric acid,mandelic acid, or lactic acid can result in formation of thediastereomeric salts.

Alternatively, by method (2), the substrate to be resolved is reactedwith one enantiomer of a chiral compound to form a diastereomeric pair(Eliel, E. and Wilen, S. (1994) Stereochemistry of Organic Compounds,John Wiley & Sons, Inc., p. 322). Diastereomeric compounds can be formedby reacting asymmetric compounds with enantiomerically pure chiralderivatizing reagents, such as, for example, menthyl derivatives,followed by separation of the diastereomers and hydrolysis to yield thefree, enantiomerically enriched substrate. A method of determiningoptical purity involves making chiral esters, such as, for example, amenthyl ester, e.g., (−) menthyl chloroformate in the presence of base,or Mosher ester, α-methoxy-α-(trifluoromethyl)phenyl acetate (Jacob III.(1982) J. Org. Chem. 47:4165), of the racemic mixture, and analyzing theNMR spectrum for the presence of the two atropisomeric diastereomers.Stable diastereomers of atropisomeric compounds can be separated andisolated by normal- and reverse-phase chromatography following methodsfor separation of atropisomeric naphthyl-isoquinolines (Hoye, T., WO96/15111). By method (3), a racemic mixture of two enantiomers can beseparated by chromatography using a chiral stationary phase (ChiralLiquid Chromatography (1989) W. J. Lough, Ed. Chapman and Hall, NewYork; Okamoto, (1990) J. of Chromatogr. 513:375-378). Enriched orpurified enantiomers can be distinguished by methods used to distinguishother chiral molecules with asymmetric carbon atoms, such as, forexample, optical rotation and circular dichroism.

Schemes and Examples

General aspects of these exemplary methods are described below and inthe Examples. Each of the products of the following processes isoptionally separated, isolated, and/or purified prior to its use insubsequent processes.

A number of exemplary methods for the preparation of compounds of thedisclosure are provided herein, for example, in the Examples below.These methods are intended to illustrate the nature of such preparationsand are not intended to limit the scope of applicable methods. Certaincompounds of the disclosure can be used as intermediates for thepreparation of other compounds of the disclosure. In the exemplarymethods described herein, the fragment E-V— can also be written as R9-.PG represents a protecting group common for the given functional groupthat it is attached. The installation and removal of the protectinggroup can be accomplished using standard techniques, such as thosedescribed in Wuts, P. G. M., Greene, T. Protective Groups in OrganicSynthesis, 4th ed.; John Wiley & Sons, Inc.: Hoboken, N.J., 2007.

Scheme 1 shows a general synthesis of an E-V—C(═O)—P—W—P—C(═O)—V—Emolecule of the disclosure wherein, for illustrative purposes, E ismethoxycarbonylamino. The treatment of either 1a or 1c with one or twoequivalents respectively of methyl chloroformate under basic conditions(e.g. sodium hydroxide) provides the molecule 1b or 1d.

Scheme 2 shows a general synthesis of an E-V—C(═O)—P—W—P—C(═O)—V—Emolecule of the disclosure wherein, for illustrative purposes, P ispyrrolidine. Coupling of amine 2a with acid 2b is accomplished using apeptide coupling reagent (e.g. HATU) to afford 2c. Alternatively, amine2d is coupled with two equivalents of 2b under similar conditions toprovide 2e. Alternatively, amine 2d is reacted with two equivalents of2b′ directly to provide 2e where E′ is a leaving group such ashydroxybenztriazole, para-nitrophenol or the like making the structure2b′ an activated ester.

Scheme 3 shows a general synthesis of an R¹—V—C(═O)—P—R² intermediatewherein, for illustrative purposes, P is pyrrolidine, R¹ is a genericgroup that is depicted as either -E or a amino protecting group, and R²is a generic group that is depicted as —W—P—C(═O)—V—E,—W—P—C(═O)—V—NH-PG, —W—P—NH-PG, or —W—NH-PG. Coupling of amine 3a (or3d, 3h, 3k) with acid 3b or 3e is accomplished using a peptide couplingreagent (e.g. HATU) to afford 3c (or 3f, 3g, 3i, 3j, 3l, 3m)respectively.

Scheme 4 shows a general synthesis of an E-V—C(═O)—R¹ intermediatewherein, for illustrative purposes, E is methoxycarbonylamino and R¹ isa generic group that is depicted as either —P—W—P—C(═O)—V—NH-PG,—P—W—P-PG, —P—W-PG, —P-PG, or —O-PG. Treatment of 4a (or 4c, 4e, 4g, 4i)with methyl chloroformate under basic conditions (e.g. sodium hydroxide)provides the molecule 4b (or 4d, 4f, 4h, 4j).

Scheme 5 shows a general synthesis of an R¹—P—W—P—R² intermediate of thedisclosure wherein, for illustrative purposes, R¹ and R² are independentprotecting groups and W is a two aromatic ring unit constructed via atransition metal mediated cyclization. Alkylation of phenol 5b with analkyl bromide, such as 5a, provides the ether 5c. Cyclization of thearomatic rings in the presence of a palladium catalyst provides thecompound 5d. Treatment of 5d with CuBr₂ provides the α-haloketone 5e,which provides 5f upon addition of an acid under basic conditions (e.g.Et₃N). Reaction of 5f with an amine or amine salt (e.g. ammoniumacetate) affords the imidazole containing molecule 5g. Oxidation of 5g,5i, or 5l can be accomplished by heating in the presence of MnO₂ toprovide 5h, 5j, or 5m, respectively. Conversion of 5g or 5h with apalladium catalyst, such as Pd₂dba₃ and X-Phos, and a boron source suchas bis(pinacolato)diboron provides the boronic ester 5i or 5j. Theboronic ester is coupled with an appropriate coupling partner (e.g. 5k)using a palladium catalyst, such as Pd(PPh₃)₄ or PdCl₂(dppf), to afford5l or 5m. For each transition metal mediated cross-coupling reaction,the roles of the nucleophile and electrophile can be reversed to providethe same coupling product. Other transition metal mediated crosscouplings that enable the construction of W, but employ alternativecoupling partners and reagents, include, but are not limited to, theNegishi, Kumada, Stille, and Ullman couplings. For the preparation ofalternate two aromatic ring containing W groups, this general scheme canbe applied through the appropriate choice of the starting reagents.

Scheme 6 shows a general synthesis of an R¹—P—W—P—R² intermediate of thedisclosure wherein, for illustrative purposes, R¹ and R² are independentprotecting groups and W is a two aromatic ring unit constructed via atransition metal mediated cyclization. Treatment of 5d with an activatedvinyl reagent (e.g. potassium vinyltrifluoroborate) in the presence of apalladium catalyst (e.g. palladium acetate and S-Phos) provides thevinyl compound 6a. Conversion to the corresponding α-halo ketone can beaccomplished by bromination with N-bromosuccinimide, followed byoxidation with MnO₂. Displacement of the α-halo ketone proceeds by theaddition of an acid under basic conditions (e.g. Et₃N). Bromination of6d proceeds upon treatment with pyridinium tribromide, and is followedby the addition of a second acid under basic conditions to provide thediester 6e. Reaction of 6e with an amine or amine salt (e.g. ammoniumacetate) affords the imidazole containing molecule 6f. Oxidation of 6fcan be accomplished in the presence of MnO₂ to provide 6g.

Scheme 7 shows a general synthesis of an E-V—C(═O)—P—W—P—R intermediateof the disclosure wherein, for illustrative purposes, R is a protectinggroup and W is a two aromatic ring unit. Displacement of the α-haloketone 6b proceeds by the addition of an acid under basic conditions(e.g. Et₃N). Bromination of 7b proceeds upon treatment with pyridiniumtribromide, and is followed by the addition of a second acid under basicconditions to provide the diester 7c. Reaction of 7c with an amine oramine salt (e g ammonium acetate) affords the imidazole containingmolecule 7d. Oxidation of 7d can be accomplished in the presence of MnO₂to provide 7e.

Scheme 8 shows a general synthesis of an E-V—C(═O)—P—W—P—R intermediateof the disclosure wherein, for illustrative purposes, R is a protectinggroup and W is a two aromatic ring unit. Displacement of the α-haloketone 6d proceeds by the addition of an acid under basic conditions(e.g. Et₃N). Reaction of 8a with an amine or amine salt (e.g. ammoniumacetate) affords the imidazole containing molecule 8b. Oxidation of 8bcan be accomplished in the presence of MnO₂ to provide 8c.

Scheme 9 shows a general synthesis of an E-V—C(═O)—P—W—P—C(═O)—V—Emolecule of the disclosure wherein, for illustrative purposes, E isethylcarbonylamino. The treatment of either 9a or 9c with one or twoequivalents respectively of propionyl chloride under basic conditions(e.g. sodium hydroxide) provides the molecule 9b or 9d.

Scheme 10 shows an alternate general synthesis of an R¹—P—W—P—R²intermediate of the invention wherein, for illustrative purposes, R¹ andR² are independent protecting groups and W is a two aromatic ring unitconstructed via a transition metal mediated cyclization. Bromination of6b with a brominating agent (i.e. pyridinium tribromide) provides thedibromide 10a. Displacement of the primary bromide then proceeds by theaddition of an acid under basic conditions (e.g. K₂CO₃) to provide 10d.Conversion to 10f or 10g can be accomplished following methods describedin Scheme 8.

Scheme 11 shows an alternate general synthesis of an E-V—C(═O)—P—W—P—Rintermediate of the invention wherein, for illustrative purposes, R is aprotecting group and W is a two aromatic ring unit. Bromination of 6bwith a brominating agent (i.e. pyridinium tribromide) provides thedibromide 10a. Displacement of the primary bromide then proceeds by theaddition of an acid under basic conditions (e.g. K₂CO₃) to provide 11b.Conversion to 11d or 11e can be accomplished following methods describedin Scheme 8.

Scheme 12 shows a general synthesis of an R¹—V—C(═O)—P—R² intermediatewherein, for illustrative purposes, P is pyrrolidine, R¹ is a genericgroup that is depicted as either -E or a amino protecting group, and R²is a generic group that is depicted as —C(═O)—O—PG. Coupling of amine12a (or 12d) with acid 12b or 12e is accomplished using a peptidecoupling reagent (e.g. HATU) to afford 12c (or 120 respectively. Theconversion of 12f to 12c can be accomplished by removal of theappropriate protecting group, followed by treatment with methylchloroformate under basic conditions (e.g. sodium hydroxide).

Scheme 13 shows an alternate general synthesis of anE-V—C(═O)—P—W—P—C(═O)—V—E intermediate of the invention wherein, forillustrative purposes, W is a two aromatic ring unit. Displacement ofthe both bromides proceeds by the addition of an acid under basicconditions (e.g. K₂CO₃) to provide 11c. Conversion to 11d or 11e can beaccomplished following methods described in Scheme 8.

Specific Embodiments

In one embodiment, provided is a compound of formula (I):

E^(1a)-V^(1a)—C(═O)—P^(1a)—W^(1a)—P^(1b)—C(═O)—V^(1b)-E^(1b)  (I)

wherein:

W^(1a) is

and W^(1a) is optionally substituted with one or more halo, alkyl,haloalkyl, optionally substituted aryl, optionally substitutedheterocycle, or cyano;

Y⁵ is —O—CH₂—, —CH₂—O—, —O—C(═O)—, or —C(═O)—O—;

X⁵ is —CH₂—CH₂—, or —CH═CH—;

P^(1a) and P^(1b) are each independently:

V^(1a) and V^(1b) are each independently:

provided that at least one of V^(1a) and V^(1b) is

E^(1a) and E^(1b) are each independently —N(H)(alkoxycarbonyl),—N(H)(cycloalkylcarbonyl), or —N(H)(cycloalkyloxycarbonyl); orE^(1a)-V^(1a) taken together are R^(9a); or E^(1b)-V^(1b) taken togetherare R^(9b); and

R^(9a) and R^(9b) are each independently:

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment the disclosure provides a compound which has formula:

wherein the imidazole ring shown in formula A1, A2, A3, and A4 isoptionally substituted with one or more halo, haloalkyl, cyano, oralkyl;

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment the disclosure provides a compound which has formula:

wherein the imidazole ring shown in formula A2 and A4 is optionallysubstituted with one or more halo, haloalkyl, cyano, or alkyl;

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment P^(1a) and P^(1b) are each independently:

In one embodiment V^(1a) and V^(1b) are each independently:

provided that at least one of V^(1a) and V^(1b) is

In one embodiment, provided is a compound of formula (I):

E^(1a)-V^(1a)—C(═O)—P^(1a)—W^(1a)—P^(1b)—C(═O)—V^(1b)-E^(1b)  (I)

wherein:

W^(1a) is

and W^(1a) is optionally substituted with one or more halo, alkyl,haloalkyl, or cyano;

Y⁵ is —O—CH₂—, or —CH₂—O—;

X⁵ is —CH₂—CH₂—, or —CH═CH—;

P^(1a) and P^(1b) are each independently:

V^(1a) and V^(1b) are each independently:

provided that at least one of V^(1a) and V^(1b) is

E^(1a) and E^(1b) are each independently —N(H)(alkoxycarbonyl),—N(H)(cycloalkylcarbonyl), or —N(H)(cycloalkyloxycarbonyl); orE^(1a)-V^(1a) taken together are R^(9a); or E^(1b)-V^(1b) taken togetherare R^(9b); and

R^(9a) and R^(9b) are each independently:

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, one of V^(1a) and V^(1b) is:

and the other of V^(1a) and V^(1b) is

In one embodiment, P^(1a) and P^(1b) are each independently:

In one embodiment, one of V^(1a) and V^(1b) is:

In one embodiment, both of V^(1a) and V^(1b) are:

In one embodiment, one of V^(1a) and V^(1b) is:

In one embodiment, both of V^(1a) and V^(1b) are:

In one embodiment, one of V^(1a) and V^(1b) is:

provided that bond (a) is connected to E^(1a) or E^(1b) and bond (b) isconnected to the —C(═O)— group of formula (1) or (A1, A2, A3, or A4).

In one embodiment, one of V^(1a) and V^(1b) is:

provided that bond (a) is connected to E^(1a) or E^(1b) and bond (b) isconnected to the —C(═O)— group of formula (1) or (A1, A2, A3, or A4).

In one embodiment, one of V^(1a) and V^(1b) is:

provided that bond (a) is connected to E^(1a) or E^(1b) and bond (b) isconnected to the —C(═O)— group of formula (1) or (A1, A2, A3, or A4).

In one embodiment, one of V^(1a) and V^(1b) is:

provided that bond (a) is connected to E^(1a) or E^(1b) and bond (b) isconnected to the —C(═O)— group of formula (1) or (A1, A2, A3, or A4).

In one embodiment, P^(1a) and P^(1b) are each independently:

In one embodiment, one of P^(1a) and P^(1b) is:

In one embodiment, one of P^(1a) and P^(1b) is:

In one embodiment, both of P^(1a) and P^(1b) are:

In one embodiment, —V^(1a)—C(═O)—P^(1a)— and —P^(1b)—C(═O)—V^(1b)— areeach independently:

provided that at least one of V^(1a) and V^(1b) is

In one embodiment, —V^(1a)—C(═O)—P^(1a)— and —P^(1b)—C(═O)—V^(1b)— areeach independently:

provided that at least one of V^(1a) and V^(1b) is

In one embodiment, one of —V^(1a)—C(═O)—P^(1a)— and—P^(1b)—C(═O)—V^(1b)— is:

and the other of —V^(1a)—C(═O)—P^(1a)— and —P^(1b)—C(═O)—V^(1b)— is:

In one embodiment, one of —V^(1a)—C(═O)—P^(1a)— and—P^(1b)—C(═O)—V^(1b)— is:

and the other of —V^(1a)—C(═O)—P^(1a)— and —P^(1b)—C(═O)—V^(1b)— is:

In one embodiment, both of —V^(1a)—C(═O)—P^(1a)— and—P^(1b)—C(═O)—V^(1b)— are:

In one embodiment, at least one of E^(1a) and E^(1b) is—N(H)(alkoxycarbonyl).

In one embodiment, both of E^(1a) and E^(1b) are —N(H)(alkoxycarbonyl).

In one embodiment, at least one of E^(1a) and E^(1b) is —N(H)C(═O)OMe.

In one embodiment, both of E^(1a) and E^(1b) are —N(H)C(═O)OMe.

In one embodiment, the disclosure provides a compound of formula:

or a pharmaceutically acceptable salt or prodrug thereof.

In another embodiment, the disclosure provides a compound of formula:

or a pharmaceutically acceptable salt or prodrug thereof.

In one embodiment, the disclosure provides a compound of formula:

In one embodiment, the disclosure provides a compound of formula:

The disclosure will now be illustrated by the following non-limitingExamples. The following abbreviations are used throughout thespecification, including the Examples.

% F % Bioavailability (g) Gas ° C. Degree Celsius Ac Acetate ACNAcetonitrile approx./apprx. Approximate AUC Area under the curve BnBenzyl BOC/Boc tert-Butoxycarbonyl br Broad calc'd Calculated CC₅₀ 50%Cytotoxicity concentration d Doublet dba dibenzalacetone DCMDichloromethane dd Doublet of doublets DIPEA/DIEAN,N-Diisopropylethylamine DMA N,N-Dimethylacetamide DMAP4-Dimethylaminopyridine DMEM Eagle's minimal essential medium DMFDimethylformamide DMSO/dmso Dimethylsulfoxide dppf1,1′-bis(diphenylphosphanyl) ferrocene EC₅₀ Half maximal effectiveconcentration EDTA Ethylenediaminetetraacetic acid ESI Electrosprayionization Et Ethyl FBS Fetal bovine serum g Gram HATU2-(1H-7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uroniumhexafluorophosphate Methanaminium HPLC High performance liquidchromatography hr/h Hour Hz Hertz i.d. Inner diameter IPAmIsopropylamine IV Intravenous J Coupling constant L Liter LCMS Liquidchromatography mass spectrometry M Molar m Multiplet m/z Mass to chargeM+ Mass peak Me Methyl mg Milligram MHz Megahertz min Minute mLMilliliter mL Milliliter mM Millimolar mm Millimeter mmol Millimole MSMass spectrometry MTBE Methyl tert-butyl ether N Normal NADPHNicotinamide adenine dinucleotide phosphate NBS N-Bromosuccinimide nmNanometer NMR Nuclear magnetic resonance o/n Over night Papp Apparentpermeability PBS Phosphate buffer system Pd/C Palladium on carbon PEGPolyethylene glycol Ph Phenyl Piv Pivalate Py/pyr Pyridine q Quartetquant Quantitative rt/RT Room temperature s Singlet SFC Supercriticalfluid chromatography SPhos/S-Phos2-Dicyclohexylphosphino-2′,6′-dimethoxybiphenyl SRM Selected reactionmonitoring t Triplet t-Bu tert-Butyl TEA Triethylamine TEMPO(2,2,6,6-Tetramethyl-piperidin-1-yl)oxyl Tf TrifluoromethanesulfonateTFA Trifluoroacetic acid THF Tetrahydrofuran TLC Thin layerchromatography TMS Trimethylsilyl UV Ultraviolet w/w Weight to weightX-Phos/XPhos 2-Dicyclohexylphosphino-2′,4′,6′- triisopropylbiphenyl δChemical shift μL Microliter μm Micromolar

EXAMPLES Intermediate 1

7-(2-Bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1(2H)-one

To a stirred solution of 7-hydroxy-1-tetralone (13.9 g, 85.7 mmol) and1-bromo-2-(bromomethyl)-4-chlorobenzene (25.6 g, 90.0 mmol) indimethylformamide (850 mL) was added potassium carbonate (24 g, 172mmol). The reaction was stirred under argon for 18 hours then dilutedwith ethyl acetate (1 L). The organics were washed three times withwater and once with brine. The organic layer was then dried withmagnesium sulfate, filtered and concentrated. To the resulting oil wasadded methanol (500 mL) and the suspension was agitated for thirtyminutes. 7-(2-bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1(2H)-one(27.8 g, 89% yield) was isolated by filtration.

3-Chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

To a 1 L flask containing palladium(II) pivalate (1.18 g, 3.8 mmol),tri(4-fluorophenyl)phosphine (1.20 g, 3.8 mmol), pivalic acid (2.33 g,22.8 mmol) and potassium carbonate (31.8 g, 228 mmol) was added asolution of7-(2-bromo-5-chlorobenzyloxy)-3,4-dihydronaphthalen-1(2H)-one (27.8 g,76.2 mmol) in dimethyacetamide (380 mL). The flask was evacuated andbackfilled with argon 5 times and then stirred under argon at 60° C. for24 hours. The reaction was cooled to room temperature and diluted withMTBE and water. The resulting biphasic mixture was stirred for 3 hoursand filtered through Celite, rinsing with MTBE. The organic layer of thefiltrate was separated and then washed twice with water and once withbrine. The organics were then dried with magnesium sulfate, filtered,concentrated and purified by flash column chromatography (Hexanes/DCM)to yield 3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (14.4g, 67% yield) as an off-white solid.

3-Vinyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A 3-neck oven-dried 500 mL round-bottom flask was cooled under Ar, thencharged with 3-Chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(12.0 g, 42.1 mmol), potassium vinyltrifluoroborate (8.47 g, 6.32 mmol),Pd(OAc)₂ (473 mg, 2.11 mmol), SPhos (1.74 g, 4.25 mmol), K₂CO₃ (17.5 g,126 mmol) and anhydrous propanol (120 mL). The reaction mixture wassparged with Ar for 16 min, then heated to reflux for 5.5 h. Uponcompletion, the reaction mixture was cooled to RT and concentrated underreduced pressure. The crude residue was suspended in DCM, then washedwith H₂O and brine. The organic solution was dried over MgSO₄, filteredand concentrated under reduced pressure. The resulting residue wasfurther purified via silica plug, eluting with DCM to afford3-vinyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (10.2 g, 87%).

3-(2-Bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

3-Vinyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (9.98 g, 36.1mmol) was dissolved in a stirred solution of THF (70 mL), DMSO (70 mL)and H₂O (35 mL). NBS (6.75 g, 37.9 mmol) was added in a single portionand the reaction mixture was stirred at RT for 33 min. Upon completion,the reaction medium was diluted with EtOAc and washed twice with H₂O andonce with brine. The organic phase was dried over MgSO₄, filtered andconcentrated under reduced pressure. The resulting crude bromohydrin wassuspended in DCM (200 mL) and treated with activated MnO₂ (62.7 g, 722mmol). After stirring for 15 h at RT, the reaction mixture was filteredover celite and the filter cake was rinsed several times with DCM. Thecombined filtrate (˜400 mL) was treated with MeOH (˜100 mL) and themixture was gradually concentrated under reduced pressure, causing solidmaterial to precipitate from solution. When the liquid volume reached˜200 mL, the solid was filtered off and rinsed with MeOH. Theconcentration/precipitation/filtration/rinsing sequence was performed 2×more, resulting in the collection of 3 crops of powdered3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (7.49g, 56% over 2 steps).

Intermediate 2

9-Bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A mixture of3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (2.58g, 6.95 mmol), pyridinium tribromide (2.56 g, 8.0 mmol), dichloromethane(22 mL) and methanol (2.5 mL) was stirred at about 20° C. for 3 hours toobtain a slurry. The precipitated product was filtered, washed withdichloromethane (10 mL) and dried in a vacuum oven at 40° C. to give9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(2.62 g, 84% yield). 400 MHz ¹H NMR (CDCl₃) δ 8.03-8.01 (m, 1H), 7.85(d, J=8.2 Hz, 1H), 7.82 (s, 1H), 7.71 (s, 1H), 7.67 (s, 1H), 5.19 (s,2H), 4.74 (dd, J=4.1, 4.1 Hz, 1H), 4.45 (s, 2H), 3.37-3.29 (m, 1H),2.99-2.92 (m, 1H), 2.59-2.46 (m, 2H).

Intermediate 2a

9-Bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

To3-(2-bromo-1-hydroxyethyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(20.3g, 54.4 mmol) in DCM (365 mL) was added MeOH (22 mL) and pyridiniumtribromide (18.24 g, 57.0 mmol). After 2h, water was added (100 mL) andafter briefly agitating the layers split and the bottom organic layerwas collected. The organic layer was then washed with 1M HCl (100 mL)and the bottom organic layer containing9-bromo-3-(2-bromo-1-hydroxyethyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-onewas collected. 400 MHz ¹H NMR (CDCl₃) 7.75 (d, J=8.1 Hz, 1H), 7.68 (s,1H), 7.61 (s, 1H), 7.42 (d, J=7.5 Hz, 1H), 7.24 (s, 1H), 5.13 (s, 2H),4.99-4.96 (m, 1H), 4.73 (dd, J=4.1, 4.1 Hz, 1H), 3.69-3.66 (m, 1H),3.58-3.53 (m, 1H), 3.35-3.27 (m, 1H), 2.96-2.90 (m, 1H), 2.58-2.44 (m,2H), C—OH not observed.

To9-bromo-3-(2-bromo-1-hydroxyethyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(approx. 54.4 mmol) in DCM (365 mL) was added sodium bicarbonate (5.45g), sodium bromide (6.14 g), TEMPO (16.55 mg) and water (60 mL). Thesolution was cooled between 0-5° C. and 6% bleach (91.5 mL) was added.After 1 h isopropyl alcohol (20 mL) was added and the reaction mixturewas warmed to room temperature. Agitation was stopped, the layersseparated and the lower organic layer was collected and concentratedremoving approximately 345 g of solvent. The slurry was filtered and thecake washed with 50 mL water and then 50 mL DCM (pre-cooled to 5° C.).The solids were collected and dried under vacuum to obtain9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(18.6 g, 76% yield). 400 MHz ¹H NMR (CDCl₃) δ 8.03-8.01 (m, 1H), 7.85(d, J=8.2 Hz, 1H), 7.82 (s, 1H), 7.71 (s, 1H), 7.67 (s, 1H), 5.19 (s,2H), 4.74 (dd, J=4.1, 4.1 Hz, 1H), 4.45 (s, 2H), 3.37-3.29 (m, 1H),2.99-2.92 (m, 1H), 2.59-2.46 (m, 2H); 100 MHz ¹³C NMR (CDCl₃) δ 190.4,189.6, 154.2, 136.6, 134.1, 133.9, 132.9, 131.8, 129.3, 127.2, 125.6,124.2, 123.3, 117.0, 68.1, 49.9, 31.8, 30.4, 25.5.

Intermediate 2b

3-((Trimethylsilyl)ethynyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A 300 mL flask equipped with an overhead stirrer and a reflux condenserunder an atmosphere of nitrogen was charged with3-chloro-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (10.0g, 35.12mmol), powdered anhydrous tripotassium phosphate (22.4 g, 105.4 mmol),XPhos (1.34 g, 2.81 mmol), and PdCl₂(MeCN)₂ (364 mg, 1.40 mmol).Acetonitrile (140 mL) was added followed by TMS acetylene (18 mL, 141mmol). The mixture was heated to 65° C. After 6h, the reaction wasjudged complete, and the mixture was cooled to 20° C. The mixture wasfiltered through a fritted funnel, and the filter cake was washed withacetonitrile. The filtrate was concentrated to about 150 mL underreduced pressure and extracted with heptane (50 mL, 3×100 mL). N-Acetylcysteine (15 g) was added to the acetonitrile phase, and the mixture wasagitated for 5 h at 45° C. The mixture was cooled to ambienttemperature, filtered through a fritted funnel, and the filter cake waswashed with acetonitrile. The filtrate was concentrated to about 120 mLunder reduced pressure. Water (120 mL) was added and the mixture wasagitated for 40 minutes at 45° C. and then cooled to ambienttemperature. After 30 minutes the mixture was filtered through a frittedfunnel to provide3-((trimethylsilyl)ethynyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(4.07 g, 33.4% yield) as a yellow solid: 400 MHz ¹H NMR (CDCl₃) δ 7.65(d, J=8.1 Hz, 1H), 7.60 (s, 1H), 7.55 (s, 1H), 7.47 (dd, J=8.1, 1.4 Hz,1H), 7.27 (s, 1H), 5.06 (s, 2H), 2.95 (t, J=6.1 Hz, 2H), 2.67-2.59 (m,2H), 2.18-2.08 (m, 2H), 0.26 (s, 9H).

3-Acetyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A 20 mL vial with stirbar was charged with3-((trimethylsilyl)ethynyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(850 mg, 2.44 mmol) and formic acid (9.8 mL). The solution was heated to65° C. After 3 h, the reaction was judged complete. The mixture wasconcentrated under reduced pressure; the resulting residue was taken upin CH₂Cl₂ and loaded onto a prepacked 25g silica gel cartridge. Theproduct was purified by chromatography on a prepacked 80g silica gelcolumn eluting with a solvent gradient from 5% to 85% EtOAc/hexanes. Theproduct containing fractions were combined and concentrated to provide3-acetyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (616 mg, 86%):400 MHz ¹H NMR (CDCl₃) δ 8.00-7.94 (m, 1H), 7.81 (d, J=8.2 Hz, 1H), 7.77(s, 1H), 7.64 (s, 2H), 5.16 (s, 2H), 2.98 (t, J=6.1 Hz, 2H), 2.69-2.64(m, 2H), 2.63 (s, 3H), 2.21-2.09 (m, 2H).

9-Bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one

A 20 mL vial with a stirbar was charged with3-acetyl-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one (100 mg, 0.366mmol), 9:1 CH₂Cl₂/MeOH (3.4 mL) and pyridinium tribromide (246 mg, 0.769mmol). The solution was heated to 35° C. After 30 minutes, the reactionwas judged complete. The mixture was cooled to ambient temperature,diluted with EtOAc (50 mL) and sequentially washed with saturatedaqueous Na₂S₂O₃ (20 mL), 2% aqueous NaHCO₃ (20 mL), water (20 mL), andbrine (10 mL). The organic phase was dried over MgSO₄, filtered andconcentrated under reduced pressure resulting in9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(68 mg, 41%): 400 MHz ¹H NMR (CDCl₃) δ 8.03-8.01 (m, 1H), 7.85 (d, J=8.2Hz, 1H), 7.82 (s, 1H), 7.71 (s, 1H), 7.67 (s, 1H), 5.19 (s, 2H), 4.74(dd, J=4.1, 4.1 Hz, 1H), 4.45 (s, 2H), 3.37-3.29 (m, 1H), 2.99-2.92 (m,1H), 2.59-2.46 (m, 2H).

Intermediate 3

(S)-Ethyl 2-(tert-butoxycarbonylamino)-5-oxohexanoate

A solution of ethyl N-Boc (S)-pyroglutamate (20.0 g, 77.7 mmol) was inanhydrous THF (150 mL) in a two neck round bottom under argon was cooledto −40° C. Methyl-magnesium bromide solution (3.0 M in Ether, 28.5 mL,85.5 mmol) was added to the reaction mixture dropwise over 30 minutes.The reaction was stirred for 4 hrs at −40° C. then for 1 hr at 0° C. Thereaction was partitioned between ethyl acetate and saturated ammoniumchloride solution and acidified with 1 N HCl. The aqueous layer wasextracted two more times with ethylacetate. The organic layers werecombined and dried with sodium sulfate. The crude material was purifiedby column chromatography (20%-40% EtOAc/hexanes) to yield (S)-ethyl2-(tert-butoxycarbonylamino)-5-oxohexanoate as a viscous oil and wasused directly in the following step.

(S)-Ethyl 5-methyl-3,4-dihydro-2H-pyrrole-2-carboxylate

(S)-ethyl 2-(tert-butoxycarbonylamino)-5-oxohexanoate in a 1 L flask wastreated with a trifluoro acetic acid/dichloromethane solution (1:1mixture, 100 mL). Effervescence was observed and the mixture was allowedto stir for 4 hours at room temperature. After which time the volatileswere removed in vacuo to yield (S)-ethyl5-methyl-3,4-dihydro-2H-pyrrole-2-carboxylate as an oil, and useddirectly in the following step.

(2S,5S)-Ethyl 5-methylpyrrolidine-2-carboxylate

The crude imine 3 in a 1 L flask was dissolved with ethanol (400 mL) wasevacuated and charged with argon three times (3×). Palladium on carbon(approx. 750 mg, 10% w/w, dry) was added and the reaction was evacuatedof gas and charged with hydrogen gas (3×). The reaction was allowed tostir under atmospheric hydrogen for 16 hours. The mixture was filteredthrough a plug of celite and the filtrate was concentrated in vacuo.Diethyl ether was added to the oil and a precipitate formed. The mixturewas filtered to yield (2S,5S)-ethyl 5-methylpyrrolidine-2-carboxylate,as a white solid (10.6 g, 67.4 mmol, 86.7% over three steps). ¹H NMR(400 MHz, cdcl₃) δ 4.48 (dd, 1H), 4.27 (q, 2H), 3.92-3.80 (m, 1H),2.52-2.36 (m, 1H), 2.32-2.13 (m, 2H), 1.75-1.60 (m, 1H), 1.51 (d, 3H),1.30 (t, 3H).

(2S,5S)-1-Tert-butyl 2-ethyl 5-methylpyrrolidine-1,2-dicarboxylate

To a solution of (2S,5S)-ethyl 5-methylpyrrolidine-2-carboxylate (7.0 g,44.5 mmol) in dichloromethane (250 mL), ditertbutylanhydride (10.7 g,49.0 mmol), diisopropylethylamine (17.1 mL, 98.0 mmol) dropwise over 10minutes, and dimethyl amino pyridine (0.27 g, 2.23 mmol) were addedsuccessively. Effervescence was observed and the mixture was allowed tostir for 16 hours at room temperature. The reaction was washed with HCl(250 mL, of 1N). The organic layer was then dried with sodium sulfate.The crude material was purified by column chromatography (5%-25%EtOAc/hexanes) to yield (2S,5S)-1-tert-butyl 2-ethyl5-methylpyrrolidine-1,2-dicarboxylate as an oil (6.46 g, 25.1 mmol,56%). LCMS-ESI⁺: calc'd for C₁₃H₂₃NO₄: 257.16 (M⁺); Found: 258.70(M+H⁺).

(2S,5S)-1-(Tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid

To a solution of (2S,5S)-1-tert-butyl 2-ethyl5-methylpyrrolidine-1,2-dicarboxylate (6.46 g, 25.1 mmol) in ethanol (20mL) was added lithium hydroxide mono hydrate (2.11 g, 50.2 mmol) anddeionized water (12 mL). The mixture was allowed to stir for 16 hoursthen partitioned between ethylacetate and a 1:1 mixture of saturatedbrine and 1N HCl. The aqueous layer was extracted an additional timewith ethyl acetate. The organic layers were combined, dried with sodiumsulfate and the solvent was removed in vacuo to yield(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid asa white solid (quant.) and was used directly in the following step.

Intermediate 4

(2S,5S)-Ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate

(2S,5S)-Ethyl 5-methylpyrrolidine-2-carboxylate-TFA (10.0 g, 39.3 mmol),(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid (6.88 g, 39.3 mmol)and HATU (14.9 g, 39.3 mmol) were combined in DMF (100 mL) and DIPEA(15.0 mL, 86.5 mmol) was added. After stirring for 1 h at RT, thereaction mixture was diluted with EtOAc. The organic phase was washedsuccessively with 10% HCl, saturated aqueous NaHCO₃ and brine, thendried over MgSO₄, filtered and concentrated under reduced pressure toafford (2S,5S)-ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate.The crude material was carried on without further purification.

(2S,5S)-1-((S)-2-(Methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid

(2S,5S)-Ethyl1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylate(39.3 mmol, assuming complete conversion from the previoustransformation) was suspended in MeOH (200 mL) and aqueous LiOH (1.0 M,100 mL, 100 mmol) was added. The reaction mixture was stirred o/n, thenconcentrated under reduced pressure to remove most of the MeOH. Theaqueous solution was washed 2× with DCM before being acidified to pH-1-2with 10% HCl. The acidic aqueous phase was then extracted 5× with EtOAc.The combined EtOAc extracts were dried over MgSO₄ filtered andconcentrated under reduced pressure to afford(2S,5S)-1-((S)-2-(Methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid (6.89 g, 56% over 2 steps).

Intermediate 5

(2S,4S)-1-Tert-butyl 2,4-dimethyl pyrrolidine-1,2,4-tricarboxylate

To a solution of (2S,4S)-1-tert-butyl 2-methyl4-cyanopyrrolidine-1,2-dicarboxylate (9.0 g, 35.4 mmol) in MeOH (196 mL)was added HCl (4M in 1,4-dioxane, 100 mL, 403 mmol). The solution wasstirred at room temperature for 16h and concentrated in vacuo. The crudeintermediate was dissolved in EtOAc (180 mL) and basified with aqueousbicarbonate (sat.). Di-tert-butyl dicarbonate (8.5 g, 38.9 mmol) wasadded and the biphasic solution was stirred at room temperature for 12h.The layers were then separated and the aqueous layer was back extractedwith EtOAc. The combined organic layers were washed with brine, driedover Na₂SO₄, and concentrated. The crude oil was purified by silica gelchromatography (15% to 40% to 100% EtOAc/Hexanes) to provide(2S,4S)-1-tert-butyl 2,4-dimethyl pyrrolidine-1,2,4-tricarboxylate (9.56g, 94%).

(3S,5S)-1-(Tert-butoxycarbonyl)-5-(methoxycarbonyl)pyrrolidine-3-carboxylicacid

To a solution of (2S,4S)-1-tert-butyl 2,4-dimethylpyrrolidine-1,2,4-tricarboxylate (9.56 g, 33.3 mmol) in THF (70 mL) at0° C. (external temperature, ice bath) was added NaOH (1N aqueous, 33mL, 33.3 mmol) dropwise over 15 min. The solution was stirred at 0° C.for 5h before acidification with HCl (1N). The solution was extractedwith EtOAc (3×). The combined organic layers were dried over Na₂SO₄ andconcentrated. The crude oil was purified by silica gel chromatography(2% to 5% to 10% MeOH/CH₂Cl₂) to provide(3S,5S)-1-(tert-butoxycarbonyl)-5-(methoxycarbonyl)pyrrolidine-3-carboxylicacid (6.38g, 70%).

(2S,4S)-1-Tert-butyl 2-methyl4-(hydroxymethyl)pyrrolidine-1,2-dicarboxylate

To a solution of(3S,5S)-1-(tert-butoxycarbonyl)-5-(methoxycarbonyl)pyrrolidine-3-carboxylicacid (6.38 g, 23.3 mmol) in THF (116 mL) at 0° C. (external temperature,ice bath) was added Et₃N (4.9 mL, 35.0 mmol) and ethyl chloroformate(2.7 mL, 28.0 mmol). The resulting solution was stirred at 0° C. for 45min, during which time a white precipitate forms. The reaction mixturewas filtered through celite and concentrated.

The crude intermediate was dissolved in THF (59 mL) and cooled to 0° C.(external temperature, ice bath). NaBH₄ (4.41 g, 116.7 mmol) in H₂O (59mL) was slowly added and the resulting solution was stirred at 0° C. for2 h. The reaction mixture was diluted with EtOAc and washed with H₂O.The aqueous layer was back extracted with EtOAc. The combined organiclayers were dried over Na₂SO₄ and concentrated. The crude oil waspurified by silica gel chromatography (42% to 69% to 100% EtOAc/Hexanes)to provide (2S,4S)-1-tert-butyl 2-methyl4-(hydroxymethyl)pyrrolidine-1,2-dicarboxylate (3.63 g, 60%).

(2S,4S)-1-Tert-butyl 2-methyl4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate

To a solution of (2S,4S)-1-tert-butyl 2-methyl4-(hydroxymethyl)pyrrolidine-1,2-dicarboxylate (2.57 g, 9.9 mmol) inCH₂Cl₂ (50 mL) was added AgOTf (4.07 g, 15.8 mmol) and2,6-di-tert-butylpyridine (4.4 mL, 19.8 mmol). The reaction mixture wascooled to 0° C. (external temperature, ice bath) and MeI (0.98 mL, 15.8mmol) was slowly added. The resulting slurry was stirred at 0° C. for1.5 h and at room temperature for 1.5 h. The slurry was diluted withCH₂Cl₂ and filtered through celite. The filtrate was concentrated todryness, dissolved in Et₂O, and washed with HCl (1N) and brine. Theaqueous layers were back extracted with Et₂O and the combined organiclayers were dried over Na₂SO₄ and concentrated. The crude oil waspurified by silica gel chromatography (10% to 75% to 100% EtOAc/Hexanes)to provide (2S,4S)-1-tert-butyl 2-methyl4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (2.11 g, 78%). ¹H-NMR:400 MHz, (CDCl₃) δ: (mixture of rotomers, major reported) 4.20 (t, 1H),3.71 (s, 3H), 3.67 (m, 1H), 3.34 (m, 2H), 3.30 (s, 3H), 3.16 (t, 1H),2.43 (m, 2H), 1.74 (m, 1H), 1.38 (s, 9H).

(2S,4S)-1-(Tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid

To a solution of (2S,4S)-1-tert-butyl 2-methyl4-(methoxymethyl)pyrrolidine-1,2-dicarboxylate (2.11 g, 7.7 mmol) in amixture of THF (38 mL) and MeOH (15 mL) was added LiOH (2.5 M aqueous,15 mL, 38.6 mmol). The resulting solution was stirred at roomtemperature for 2h, and acidified with aqueous HCl (1N). The desiredproduct was extracted with CH₂Cl₂ (4×). The combined organic layers weredried over Na₂SO₄ and concentrated to provide(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid (2.0 g, 99%). ¹H-NMR: 400 MHz, (CDCl₃) δ: (mixture of rotomers,major reported) 4.33 (t, 1H), 3.65 (m, 1H), 3.35 (m, 2H), 3.32 (s, 3H),3.16 (t, 1H), 2.45 (m, 2H), 2.12 (m, 1H), 1.46 (s, 9H).

Intermediate 6

(2S,5S)-1-((2S,3S)-2-(Methoxycarbonylamino)-3-methylpentanoyl)-5methylpyrrolidine-2-carboxylic acid

(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5methylpyrrolidine-2-carboxylic acid was synthesized in a similar manneras Intermediate 4 substituting(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid with(2S,3S)-2-(methoxycarbonyl-amino)-3-methylpentanoic acid MS (ESI) m/z301.19 [M+H]⁺.

Example 7

(2S,5S)-1-(Tert-butoxycarbonyl)-5-ethylpyrrolidine-2-carboxylic acid

(2S,5S)-1-(tert-butoxycarbonyl)-5-ethylpyrrolidine-2-carboxylic acid wassynthesized in a similar manner as Intermediate 3 substitutingethylmagnesium bromide for methylmagnesium bromide. ¹HNMR (400 MHz,DMSO-d6): δ 12.37 (1H, s), 4.05-4.07 (1H, m), 3.63-3.64 (1H, m),2.13-2.15 (1H, m), 1.63-1.90 (4H, m), 1.39 (10H, m), 0.83 (3H, t, J=7.2Hz).

Example 8

(2S,5S)-5-Ethyl-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidine-2-carboxylicacid

(2S,5S)-5-ethyl-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)pyrrolidine-2-carboxylicacid was synthesized in a similar manner as Example 4 substituting(2S,5S)-ethyl 5-methylpyrrolidine-2-carboxylate-TFA with (2S,5S)-methyl5-ethylpyrrolidine-2-carboxylate-HCl. MS (ESI) m/z 301.15 [M+H]⁺.

Intermediate 9

(2S,5S)-5-Ethyl-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)pyrrolidine-2-carboxylicacid

(2S,5S)-5-ethyl-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)pyrrolidine-2-carboxylicacid was synthesized in a similar manner as Intermediate 4 substituting(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid with(2S,3S)-2-(methoxycarbonyl-amino)-3-methylpentanoic acid and(2S,5S)-ethyl 5-methylpyrrolidine-2-carboxylate-TFA with (2S,5S)-methyl5-ethylpyrrolidine-2-carboxylate-HCl.

Intermediate 10

(S)-1-Tert-butyl 2-methyl 4-oxopyrrolidine-1,2-dicarboxylate

CrO₃ (194 g, 1.94 mol) was added slowly with stirring over 30 min to asolution of pyridine (340 mL) in DCM (900 mL) at 0° C. The mixture waswarmed to rt and (2S,4R)-1-tert-butyl 2-methyl4-hydroxypyrrolidine-1,2-dicarboxylate (56 g, 0.216 mol) in DCM (700 mL)was added. The reaction was stirred vigorously for 4 hrs at rt. Theformed dark solid was decanted and washed with DCM. The organic phaseswere washed with aq. NaHCO₃, 10% aqueous critic acid, and brine, anddried over anhydrous Na₂SO₄. The solvent was removed in vacuo andpurified by silica gel column chromatography (PE:EtOAc=50:1 to 10:1) toafford (S)-1-tert-butyl 2-methyl 4-oxopyrrolidine-1,2-dicarboxylate(42.6 g, 81%) as yellow oil.

(S)-1-Tert-butyl 2-methyl 4-ethylidenepyrrolidine-1,2-dicarboxylate

A solution of Ph₃PEtBr (84 g, 227 mmol) and KOtBu (76.7 g, 556 mmol) inTHF (1100 mL) was stirred at rt under nitrogen atmosphere for 1 h, andthen added (S)-1-tert-butyl 2-methyl 4-oxopyrrolidine-1,2-dicarboxylate(50 g, 206 mmol) in THF (350 mL) dropwise. The mixture was stirred atroom temperature for 4 hrs. TLC showed the reaction was completed. Themixture was quenched with NH4Cl aqueous and concentrated to remove THF,and then dissolved in EtOAc and water. The combined organic layer waswashed with water, brine, dried over Na2SO4, filtered and concentrated.The crude product was purified by column chromatography (PE:EtOAc=30:1to 5:1) to afford (S)-1-tert-butyl 2-methyl4-ethylidenepyrrolidine-1,2-dicarboxylate (18.3 g, 35%) as yellow oil.

(2S)-1-Tert-butyl 2-methyl 4-ethylpyrrolidine-1,2-dicarboxylate

A mixture of (S)-1-tert-butyl 2-methyl4-ethylidenepyrrolidine-1,2-dicarboxylate (50 g, 196 mmol), Pd/C (5 g)in EtOH (500 mL) was hydrogenated at room temperature overnight. Themixture was filtered and concentrated to afford (2S)-1-tert-butyl2-methyl 4-ethylpyrrolidine-1,2-dicarboxylate (9.8 g, 97%) as colorlessoil.

(2S)-1-(Tert-butoxycarbonyl)-4-ethylpyrrolidine-2-carboxylic acid

A mixture of (2S)-1-tert-butyl 2-methyl4-ethylpyrrolidine-1,2-dicarboxylate (49.5 g, 0.19 mol), LiOH (950 mL,1M) in MeOH (1500 mL) was stirred at room temperature overnight. TLCshowed the reaction was completed. The mixture was concentrated,adjusted the pH to 2 with 1N HCl. The mixture was extracted with EA, thecombined organic layer was washed with brine, dried over Na₂SO₄,concentrated to afford(2S)-1-(tert-butoxycarbonyl)-4-ethylpyrrolidine-2-carboxylic acid (45.5g, 97%) as white solid without further purification.

(2S)-2-Benzyl 1-tert-butyl 4-ethylpyrrolidine-1,2-dicarboxylate

A mixture of(2S)-1-(tert-butoxycarbonyl)-4-ethylpyrrolidine-2-carboxylic acid (45.5g, 187 mmol), TEA (37.8 g, 374 mmol) in THF (1 L) was added dropwiseBnBr (38.5 g, 225 mmol) at 0° C. The mixture was stirred at roomtemperature overnight. TLC showed the reaction was completed. Themixture was concentrated to remove solvent. The residue was partitionedbetween EtOAc and water. The combined organic layer was washed withbrine, dried over Na₂SO₄ and concentrated. The crude product waspurified by column chromatography to give (2S)-2-benzyl 1-tert-butyl4-ethylpyrrolidine-1,2-dicarboxylate (46 g, 74%) as colorless oil.(2S)-2-benzyl 1-tert-butyl 4-ethylpyrrolidine-1,2-dicarboxylate wasseparated by preparative SFC via a Chiralcel OD 250*50 mm i.d. 10 μmcolumn (Mobile phase: A for n-hexane and B for ethanol (0.05% IPAm),Gradient: A:B=97:3, Flow rate: 100 ml/min, Wavelength: 210 and 220 nm,Injection amount: 0.4 g per injection) to provide (2S,4S)-2-benzyl1-tert-butyl 4-ethylpyrrolidine-1,2-dicarboxylate.

(2S,4S)-1-(Tert-butoxycarbonyl)-4-ethylpyrrolidine-2-carboxylic acid

A mixture of (2S,4S)-2-benzyl 1-tert-butyl4-ethylpyrrolidine-1,2-dicarboxylate (18 g, 54.1 mmol), Pd/C (3.6 g) inMeOH (1 L) was hydrogenated at room temperature overnight. TLC showedthat the reaction was completed. The mixture was filtered by Celite. Thefiltrate was concentrated to afford(2S,4S)-1-(tert-butoxycarbonyl)-4-ethylpyrrolidine-2-carboxylic acid (10g, 77%) as white solid. ¹H NMR: 400 MHz CDCl₃: δ 9.88 (br, 1H),4.31-4.19 (m, 1H), 3.82-3.68 (m, 1H), 3.03-2.95 (m, 1H), 2.49-2.39 (m,1H), 2.12-2.03 (m, 1H), 1.81-1.56 (m, 1H), 1.45 (d, J=8 Hz, 11H), 0.92(t, J=6 Hz, 3H).

Intermediate 11

rel-(2S,4S,5S)-1-(tert-butoxycarbonyl)-4,5-dimethylpyrrolidine-2-carboxylicacid

To a solution of 1-tert-butyl 2-ethyl4,5-dimethyl-1H-pyrrole-1,2-dicarboxylate (4.016 g, 15.02 mmol) in EtOH(100 mL) was added Platinum on carbon (5%, 0.58 g). The slurry wasstirred under an atmosphere of hydrogen (1 atm) for 3 days. The slurrywas filtered through celite and washed with MeOH. The filtrate wasconcentrated and the crude was purified by column chromatography (SiO₂,5-10-20% EtOAc/Hexanes) to provide rel-(2S,4S,5S)-1-tert-butyl 2-ethyl4,5-dimethylpyrrolidine-1,2-dicarboxylate.

To a solution of rel-(2S,4S,5S)-1-tert-butyl 2-ethyl4,5-dimethylpyrrolidine-1,2-dicarboxylate in a mixture of THF (70 mL),MeOH (25 mL), and H₂O (25 mL) was added lithium hydroxide (1.53 g, 63.7mmol). The slurry was stirred at room temperature for 2.5h and at 45° C.for 2 h. The solution was cooled to room temperature and HCl (aqueous,1N, 70 mL) was added. The organics were concentrated and the resultingaqueous layer was extracted with EtOAc (3×). The combined organic layerswere dried over Na₂SO₄ and concentrated to providerel-(2S,4S,5S)-1-(tert-butoxycarbonyl)-4,5-dimethylpyrrolidine-2-carboxylicacid (3.08 g, 84%).

Intermediate 12

(2S,3aS,6aS)-2-benzyl 1-tert-butylhexahydrocyclopenta[b]pyrrole-1,2(2H)-dicarboxylate

To a solution of commercially available (2S,3aS,6aS)-2-benzyl1-tert-butyl hexahydrocyclopenta[b]pyrrole-1,2(2H)-dicarboxylate (4.70g, 16.68 mmol) in methylene chloride (42 mL) was added Di-tert-butyldicarbonate (7.28 g, 33.36 mmol) N,N-diisopropylethylamine (5.82 mL,33.36 mmol) and 4-(Dimethylamino)pyridine (0.20 g, 1.67 mmol). Thesolution was stirred under air for 16 hours. Upon completion, thereaction was concentrated in vacuo, diluted in ethyl acetate, and washedwith 1N HCl. The aqueous layers were back extracted twice with ethylacetate and the combined organic layers were dried over sodium sulfate,filtered and concentrated. The resulting residue was purified by silicagel chromatography (5-40% ethyl acetate in hexanes) to afford(2S,3aS,6aS)-2-benzyl 1-tert-butylhexahydrocyclopenta[b]pyrrole-1,2(2H)-dicarboxylate which was usedwithout further purification. MS (ESI) m/z 368.47 [M+Na]⁺.

(2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid

To a 250 mL round bottom flask charged with a stirbar and(2S,3aS,6aS)-2-benzyl 1-tert-butylhexahydrocyclopenta[b]pyrrole-1,2(2H)-dicarboxylate (5.76 g, 16.68 mmol)was added 10% Palladium on carbon (1.77g). Ethanol was poured over themixture and the reaction mixture was evacuated and flushed with hydrogengas three times. The suspension was stirred at room temperature underand atmosphere of hydrogen for 24 hours. Upon completion, the reactionmixture was filtered through celite and concentrated to give(2S,3aS,6aS)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid (4.45g, >99%). MS (ESI) m/z 256.21 [M+H]⁺.

Intermediate 13

(2S,3aS,6aS)-benzyl1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylate

To a solution of commercially available (2S,3aS,6aS)-2-benzyl1-tert-butyl hexahydrocyclopenta[b]pyrrole-1,2(2H)-dicarboxylate (10.0g,35.489 mmol) in methylene chloride (100 mL) was added(2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoic acid (10.072g, 53.23mmol), HATU (21.59g, 56.78 mmol), and DIPEA (18.59 mL, 106.46 mmol). Thereaction was stirred overnight, at which time it was concentrated invacuo, diluted in ethyl acetate and washed with HCl (1N). The aqueouslayer was back extracted with ethyl acetate, and the combined organicswere dried over sodium sulphate, filtered and concentrated. Theresulting oil was diluted in a small amount of chloroform and filteredto remove tetramethyl urea precipitate. The resulting oil was purifiedby normal phase chromatography (50% ethyl acetate in hexanes) to give(2S,3aS,6aS)-benzyl1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylate(19.53g, >99% yield) which was used without further purification.LCMS-ESI+calc'd for C23H33N2O5:417.23; Found: 417.37.

(2S,3aS,6aS)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid

To a 250 mL round bottom flask charged with a stirbar and(2S,3aS,6aS)-benzyl1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylate(19.53g crude, assumed 35.49 mmol) was added 10% Palladium on carbon(3.55g). Ethanol was poured over the mixture and the reaction mixturewas evacuated and flushed with hydrogen gas three times. The suspensionwas stirred at room temperature under and atmosphere of hydrogen for 3days. Upon completion, the reaction mixture was filtered through celiteand concentrated to give(2S,3aS,6aS)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid (13.65g, >99%). LCMS-ESI+calc'd for C16H26N2O5:327.18; Found:327.13.

Intermediate 14

(2S,3aS,6aS)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid

(2S,3aS,6aS)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid was synthesized in a similar manner as(2S,3aS,6aS)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid substituting (2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoicacid with (S)-2-(methoxycarbonylamino)-3-methylbutanoic acid.LCMS-ESI+calc'd for C15H25N2O5:313.17; Found: 313.12.

Intermediate 15

(2S,5S)-1-((S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)acetyl)-5-methylpyrrolidine-2-carboxylicacid

(2S,5S)-1-((S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)acetyl)-5-methylpyrrolidine-2-carboxylicacid was synthesized in a similar manner as Intermediate 4 substituting(S)-2-(methoxycarbonylamino)-3-methylbutanoic acid with(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid. 1H NMR (400 MHz, Chloroform-d) δ 5.33-5.16 (m, 1H), 4.70-4.59 (m,1H), 4.54 (t, 1H), 4.34-4.19 (m, 2H), 4.12 (q, 1H), 3.78-3.70 (m, 1H),3.67 (s, 3H), 2.37-2.17 (m, 3H), 2.15-2.07 (m, 1H), 2.04 (s, 1H),1.84-1.73 (m, 1H), 1.82-1.43 (m, 3H), 1.32 (d, 3H), 1.26 (d, 4H), 1.11(d, 3H), 0.96 (q, 1H). LCMS-ESI+calc'd for C17H29N2O6:357.19; Found:357.08.

Example AA

(2S,5S)-2-(9-Bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylate

To a slurry of9-bromo-3-(2-bromoacetyl)-10,11-dihydro-5H-dibenzo[c,g]chromen-8(9H)-one(4.00 g, 8.88 mmol) in dichloromethane (50 mL) was added(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid (2.80 g, 9.32 mmol) and K₂CO₃ (1.84 g, 13.31 mmol). The resultingslurry was stirred at room temperature for 18 h. The reaction wasdiluted with dichloromethane and washed with aqueous HCl (0.5 M) andBrine. The aqueous layers were back extracted with dichloromethane (2×),and the combined organic layers were dried over Na₂SO₄ and concentrated.The crude product was taken directly into the next reaction.

(2S,5S)-1-Tert-butyl2-(3-(2-((2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carbonyloxy)acetyl)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)5-methylpyrrolidine-1,2-dicarboxylate

To a solution of(2S,5S)-2-(9-bromo-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-3-yl)-2-oxoethyl1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylate(5.95 g, 8.88 mmol) in THF (60 mL) was added(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid(3.05 g, 13.3 mmol) and Cs₂CO₃ (2.31 g, 7.09 mmol). The resultingsolution was heated to 50° C. for 18 h. The solution was cooled to roomtemperature and diluted with EtOAc and washed with aqueous HCl (0.5 M).The aqueous layer was back extracted with EtOAc (2×), and the combinedorganic layers were dried over Na₂SO₄ and concentrated. The crude oilwas purified by column chromatography (SiO₂, 25-100% EtOAc (5%MeOH)/Hexanes) to provide (2S,5S)-1-tert-butyl2-(3-(2-((2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carbonyloxy)acetyl)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)5-methylpyrrolidine-1,2-dicarboxylate (3.116, 43% over 2 steps) as aorange foam. LCMS-ESI+: calc'd for C44H55N3O12:817.38 (M+); Found:817.65 (M+).

Tert-butyl(2S,5S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidine-1-carboxylate

To a solution of (2S,5S)-1-tert-butyl2-(3-(2-((2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carbonyloxy)acetyl)-8-oxo-8,9,10,11-tetrahydro-5H-dibenzo[c,g]chromen-9-yl)5-methylpyrrolidine-1,2-dicarboxylate (3.116 g, 3.57 mmol) in toluene(35 mL) was added hexamethdisilazane (6.0 mL, 28.7 mmol), and propionicacid (8.0 mL, 107.1 mmol). The solution was heated to 90° C. for 18h andcooled to room temperature. The solution was diluted with MeOH andbasified with a 1:1 mixture of NH₄OH and water. The slurry was extractedwith dichloromethane (3×). The combined organic layers were dried overNa₂SO₄ and concentrated. The crude oil was used directly in the nextstep. LCMS-ESI+: calc'd for C44H55N7O6:777.42 (M+); Found: 778.30(M+H+).

Tert-butyl(2S,5S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidine-1-carboxylate

To a solution of tert-butyl(2S,5S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidine-1-carboxylate(2.77 g, 3.5 mmol) in dichloromethane (25 mL) was added MnO₂ (9.00 g,103 mmol). The resulting slurry was stirred at room temperature for 20h. The solution was diluted with dichloromethane, filtered throughcelite, and concentrated. The crude oil was purified by columnchromatography (SiO₂, 0-5-10% EtOAc/MeOH) to provide tert-butyl(2S,5S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidine-1-carboxylate(1.10 g, 40% over 2 steps) as a brown foam. LCMS-ESI+: calc'd forC44H53N7O6:775.41 (M+); Found: 776.37 (M+H+).

Methyl{(2S,3S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate

To a solution of tert-butyl(2S,5S)-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-isoleucyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidine-1-carboxylate(0.30 g, 0.39 mmol) in a mixture of dichloromethane (4 mL) and methanol(0.5 mL) was added HCl (4M in dioxanes, 1.45 mL, 5.80 mmol). Thesolution was heated to 40° C. for 1 h and cooled to room temperature.The solution was then concentrated in vacuo. The resulting solid wasdissolved in DMF (3 mL), followed by the addition of(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid (0.11 g, 0.46 mmol), HATU (0.18 g, 0.48 mmol), anddiisopropylethylamine (0.3 mL, 1.72 mmol). The resulting solution wasstirred at room temperature for 3 h. Aqueous HCl (6M, 4 drops) was addedand the solution was purified by reverse phase HPLC (Gemini column,10-53% MeCN/H₂O/0.1% TFA). The desired fractions were combined, and theorganics were concentrated in vacuo. The resulting aqueous solution wasbasified with saturated NaHCO₃ to provide a white precipitate. The solidwas filtered and dried to provide methyl{(2S,3S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate(0.082 g, 23%) as a white powder. LCMS-ESI+: calc'd forC50H62N8O9:903.08 (M+); Found: 903.84 (M+H+). 1H NMR (400 MHz,Methanol-d4) δ 8.36-8.22 (m, 1H), 7.98-7.82 (m, 1H), 7.69-7.20 (m, 8H),5.22-5.07 (m, 3H), 5.02 (d, 1H), 4.73-4.64 (m, 1H), 4.49 (s, 3H),4.26-3.97 (m, 4H), 3.79 (d, 1H), 3.72 (s, 1H), 3.63-3.52 (m, 4H),3.47-3.32 (m, 1H), 2.61-2.43 (m, 1H), 2.32-1.98 (m, 4H), 1.95-1.78 (m,1H), 1.79-1.65 (m, 1H), 1.54 (s, 4H), 1.41 (d, 2H), 1.28-1.09 (m, 3H),1.04 (dd, 6H), 0.90 (d, 1H), 0.88-0.59 (m, 10H).

Example AB

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Following Example AA, substituting(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid for(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid and(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid for(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid,provided methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate(0.25 g). LCMS-ESI+: calc'd for C50H62N8O9:918.46 (M+); Found: 919.97(M+H+). 1H NMR (400 MHz, Methanol-d4) δ 8.39-8.11 (m, 1H), 8.09-7.40 (m,5H), 7.28 (s, 2H), 5.27-4.91 (m, 5H), 4.49 (s, 3H), 4.34-4.14 (m, 2H),4.14-4.00 (m, 3H), 3.80 (s, 1H), 3.59 (s, 2H), 3.57 (s, 3H), 3.54-3.39(m, 3H), 3.31 (s, 3H), 2.68-2.46 (m, 2H), 2.45-2.30 (m, 1H), 2.31-1.93(m, 5H), 1.94-1.81 (m, 1H), 1.50 (d, 2H), 1.46-1.25 (m, 4H), 1.10 (dd,3H), 1.06-0.90 (m, 9H), 0.87 (d, 2H).

Example AC

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-ethyl-5-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxoethyl}carbamate

Following Example AA, substituting(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid for(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid and(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid for (2S,5S)-1-(tert-butoxycarbonyl)-5-ethylpyrrolidine-2-carboxylicacid, provided methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-ethyl-5-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxoethyl}carbamate(0.21 g). LCMS-ESI+: calc'd for C50H62N8O8:902.47 (M+);

Found: 904.14 (M+H+). 1H NMR (400 MHz, Methanol-d4) δ 8.33-8.04 (m, 1H),8.01-7.68 (m, 1H), 7.68-7.37 (m, 14H), 7.32-7.17 (m, 1H), 5.22-4.95 (m,3H), 4.50 (s, 6H), 4.31-3.92 (m, 5H), 3.80 (s, 1H), 3.58 (d, J=2.9 Hz,4H), 3.52-3.35 (m, 1H), 2.56-2.39 (m, 1H), 2.31-2.11 (m, 2H), 2.12-1.88(m, 3H), 1.88-1.76 (m, 1H), 1.76-1.48 (m, 1H), 1.40 (d, J=6.7 Hz, 2H),1.21 (d, J=7.0 Hz, 2H), 1.12-1.02 (m, 4H), 0.99 (t, J=7.3 Hz, 1H), 0.93(d, J=7.0 Hz, 1H), 0.89 (d, J=6.6 Hz, 1H), 0.87-0.74 (m, 5H), 0.70-0.45(m, 1H).

Example AD

Methyl[(2S)-1-{(2S,5S)-2-[5-(2-{(2S,5S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate

Methyl[(2S)-1-{(2S,5S)-2-[5-(2-{(2S,5S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid and substituting(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(S)-2-(tert-butoxycarbonylamino)-2-(4-methyltetrahydro-2H-pyran-4-yl)aceticacid. MS (ESI) m/z 917.62 [M+H]⁺.

Methyl[(2S)-1-{(2S,5S)-2-[5-(2-{(2S,5S)-1-[(2S)-2-[(methoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate

To as solution of methyl[(2S)-1-{(2S,5S)-2-[5-(2-{(2S,5S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate(0.594g, 0.648 mmol) in a mixture of dichloromethane (6.4 mL) andmethanol (1.2 mL) was added HCl (4M in dioxanes, 2.4 mL, 9.72 mmol). Thesolution was heated to 40° C. for 1 h and cooled to room temperature.The solution was then concentrated in vacuo. The resulting solid wasdissolved in DMF (3 mL), followed by the addition of methylchloroformate (0.050 mL, 0.648 mmol) and diisopropylethylamine (0.14 mL,0.78 mmol). The reaction mixture was stirred at room temperature for 30minutes. Upon completion by LCMS monitoring, the solution was purifiedby reverse phase HPLC (Gemini column, 10-45% MeCN/H₂O/0.1% TFA). Thedesired fractions were lyophilized to give methyl[(2S)-1-{(2S,5S)-2-[5-(2-{(2S,5S)-1-[(2S)-2-[(methoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate(0.057 g, 10%). ¹H NMR (400 MHz, Methanol-d4) δ 8.63 (s, 1H), 8.19 (d,1H), 8.09-7.75 (m, 4H), 7.75-7.61 (m, 2H), 7.46-7.24 (m, 1H), 5.39-5.24(m, 2H), 5.23-5.12 (m, 1H), 4.84-4.60 (m, 2H), 4.44-4.22 (m, 1H),4.22-4.02 (m, 1H), 3.85-3.62 (m, 8H), 3.60-3.45 (m, 2H), 2.84-2.44 (m,2H), 2.42-2.23 (m, 3H), 2.12-1.82 (m, 3H), 1.67 (d, 2H), 1.56 (d, 4H),1.37-1.19 (m, 4H), 1.17-1.03 (m, 4H), 0.98 (d, 3H), 0.88 (d, 3H). MS(ESI) m/z 875.53 [M+H]⁺.

Example AE

Methyl[(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-{(2S,4S)-4-ethyl-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidin-1-yl}-2-oxoethyl]carbamate

Methyl[(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-{(2S,4S)-4-ethyl-2-[5-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]pyrrolidin-1-yl}-2-oxoethyl]carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,4S)-1-(tert-butoxycarbonyl)-4-ethylpyrrolidine-2-carboxylic acid andsubstituting(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid. 1H NMR (400 MHz, Methanol-d4) δ 8.48-8.29 (m, 1H), 8.10-7.91 (m,1H), 7.86-7.23 (m, 5H), 5.35-5.15 (m, 2H), 5.07 (t, 1H), 4.46-4.26 (m,2H), 4.25-4.07 (m, 2H), 3.91 (s, 1H), 3.66 (d, 5H), 3.52-3.37 (m, 1H),2.74-2.41 (m, 2H), 2.40-1.89 (m, 6H), 1.75-1.34 (m, 7H), 1.33-0.75 (m,18H). MS (ESI) m/z 903.99 [M+H]⁺.

Example AF

Methyl[(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-{(2S,4S)-4-ethyl-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-2-oxoethyl]carbamate

Methyl[(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-{(2S,4S)-4-ethyl-2-[9-(2-{(2S,5S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]pyrrolidin-1-yl}-2-oxoethyl]carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid and substituting(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith (2S,4S)-1-(tert-butoxycarbonyl)-4-ethylpyrrolidine-2-carboxylicacid. 1H NMR (400 MHz, Methanol-d4) δ 8.34 (d, 1H), 7.94 (dd, 1H),7.88-7.60 (m, 3H), 7.59-7.28 (m, 2H), 5.25-5.11 (m, 3H), 4.54 (s, 1H),4.39 (t, 1H), 4.27-4.12 (m, 2H), 4.12-4.02 (m, 1H), 3.62 (d, 4H), 3.48(s, 3H), 3.13 (s, 3H), 2.68-2.45 (m, 1H), 2.38-2.19 (m, 2H), 2.19-1.83(m, 4H), 1.70-1.53 (m, 2H), 1.46 (d, 2H), 1.44-1.28 (m, 3H), 1.28-1.13(m, 1H), 1.10 (d, 3H), 1.07-0.87 (m, 12H), 0.85-0.77 (m, 1H). MS (ESI)m/z 903.88 [M+H]⁺.

Example AG

Methyl{(1S)-2-[(2S,5S)-2-ethyl-5-(9-{2-[(2S,5S)-1-{(2S)-2[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-[(3R)-tetrahydro-2H-pyran-3-yl]ethyl}carbamate

Methyl{(1S)-2-[(2S,5S)-2-ethyl-5-(9-{2-[(2S,5S)-1-{(2S)-2[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-[(3R)-tetrahydro-2H-pyran-3-yl]ethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid; (2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylicacid with (2S,5S)-1-(tert-butoxy carbonyl)-5-ethylpyrrolidine-2-carboxylic acid and(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(S)-2-(methoxycarbonylamino)-2-((R)-tetrahydro-2H-pyran-3-yl)aceticacid. ¹H NMR (400 MHz, dmso) δ 8.60 (s, 1H), 8.25-7.43 (m, 7H), 5.23 (s,2H), 5.13 (m, 1H), 5.01-4.90 (m, 1H), 4.59 (s, 1H), 4.33 (m, 2H),4.12-3.43 (m, 14H), 3.37 (m, 1H), 3.08 (m, 2H), 2.39-1.70 (m, 10H), 1.44(m, 5H), 1.12 (m, 2H), 0.92 (m, 5H), 0.73-0.54 (m, 3H) MS (ESI) m/z875.95 [M+H]⁺.

Example AH

Methyl{(1S)-2-[(2S,5S)-2-ethyl-5-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-((3S)-tetrahydro-2H-pyran-3-yl]ethyl}carbamate

Methyl{(1S)-2-{(2S,5S)-2-ethyl-5-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-[(3S)-tetrahydro-2H-pyran-3-yl]ethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid; (2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-ethylpyrrolidine-2-carboxylic acid and(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(S)-2-(methoxycarbonylamino)-2-((S)-tetrahydro-2H-pyran-3-yl)aceticacid. ¹H NMR (400 MHz, dmso) δ 8.64 (s, 1H), 8.24-7.46 (m, 8H), 5.27 (s,2H), 5.13 (s, 1H), 4.99 (s, 1H), 4.62 (s, 1H), 4.12 m, 5H), 3.67-3.23(m, 8H), 3.12 (s, 1H), 2.43-2.06 (m, 6H), 2.04-1.63 (m, 8H), 1.47 (m,4H), 1.38-1.07 (m, 3H), 0.95 (m, 6H), 0.79-0.62 (m, 3H). MS (ESI) m/z875.86 [M+H]⁺.

Example AI

Methyl{(1S)-2-[(2S,5S)-2-methyl-5-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-[(3R)-tetrahydro-2H-pyran-3-yl]ethyl}carbamate

Methyl{(1S)-2-[(2S,5S)-2-methyl-5-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-[(3R)-tetrahydro-2H-pyran-3-yl]ethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylic acidand(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(S)-2-(methoxycarbonylamino)-2-((R)-tetrahydro-2H-pyran-3-yl)aceticacid. ¹H NMR (400 MHz, dmso) δ 8.60 (s, 1H), 8.25-7.46 (m, 7H), 5.23 (s,2H), 5.11 (m, 1H), 4.96 (s, 1H), 4.64 (m, 2H), 4.16-3.58 (m, 9H),3.56-3.31 (m, 6H), 3.08 (m, 3H), 2.19 (m, 5H), 1.86 (m, 3H), 1.43 (m,7H), 1.24-0.92 (m, 3H), 0.83 (m, 3H), 0.68 (m, 3H). MS (ESI) 861.45[M+H]⁺.

Example AJ

Methyl{(1S)-2-[(2S,5S)-2-methyl-5-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-[(3S)-tetrahydro-2H-pyran-3-yl]ethyl}carbamate

Methyl{(1S)-2-[(2S,5S)-2-methyl-5-(9-{2-[(2S,5S)-1-{(2S)-2-[(methoxycarbonyl)amino]-3-methylbutanoyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)pyrrolidin-1-yl]-2-oxo-1-[(3S)-tetrahydro-2H-pyran-3-yl]ethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylic acidand(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(S)-2-(methoxycarbonylamino)-2-((S)-tetrahydro-2H-pyran-3-yl)aceticacid. ¹H NMR (400 MHz, dmso) δ 8.74-8.44 (m, 1H), 8.26-7.31 (m, 9H),5.25 (s, 2H), 5.19-5.04 (m, 1H), 5.04-4.87 (m, 1H), 4.77-4.48 (m, 1H),4.44-3.73 (m, 2H), 3.66-2.95 (m, 4H), 2.29 (s, 8H), 1.83 (s, 7H), 1.46(m, 12H), 0.85 (m, 5H), 0.72 (m, 3H). MS (ESI) m/z 861.41 [M+H]⁺.

Example AK

Methyl[(2S)-1-{(2S,5S)-2-[9-(2-{(2S,5S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate

Methyl[(2S)-1-{(2S,5S)-2-[9-(2-{(2S,5S)-1-[(2S)-2-[(tert-butoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid;(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylic acidand(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(S)-2-(tert-butoxycarbonylamino)-2-(4-methyltetrahydro-2H-pyran-4-yl)aceticacid

Example AL

Methyl[(2S)-1-{(2S,5S)-2-[9-(2-{(2S,5S)-1-[(2S)-2-[(methoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate

Tert-butyl (2S,4S)-2-[5-(2-{(2S,4S)-1-[N-(methoxycarbonyl)-L-valyl]-5-methylpyrrolidin-2-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtha[1,2-d]imidazol-9-yl)-1H-imidazol-2-yl]-5-methylpyrrolidine-1-carboxylate(316 mg, 0.34 mmol) was dissolved in EtOH (3 mL) and HCl (1 mL) wasadded. The reaction mixture was stirred for 1 h at 60° C. and thenconcentrated under reduced pressure. The crude residue was dissolved inTHF (3 mL) and CH₂Cl₂ (3 mL) treated with DIPEA (0.18 mL, 1.0 mmol) andmethyl chloroformate (0.0.03 mL, 0.38 mmol). After 1 h, the mixture wasdiluted with EtOAc and washed successively with water and brine. Theorganics were dried over Na₂SO₄, filtered and concentrated under reducedpressure. The crude residue was purified by reverse phase HPLC (Gemini,15 to 45% ACN/H₂O+0.1% TFA). to afford methyl[(2S)-1-{(2S,5S)-2-[9-(2-{(2S,5S)-1-[(2S)-2-[(methoxycarbonyl)amino]-2-(4-methyltetrahydro-2H-pyran-4-yl)acetyl]-5-methylpyrrolidin-2-yl}-1H-imidazol-5-yl)-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl]-5-methylpyrrolidin-1-yl}-3-methyl-1-oxobutan-2-yl]carbamate(100 mg, 33%). ¹H NMR (400 MHz, dmso) δ 8.67 (s, 1H), 8.26-7.47 (m, 8H),5.27 (m, 2H), 5.15 (m, 1H), 5.02-4.90 (m, 1H), 4.70 (s, 1H), 4.44 (s,1H), 4.29-3.28 (m, 16H), 2.21 (m, 5H), 1.75 (m, 3H), 1.49 (m, 6H),1.35-1.05 (m, 3H), 1.02-0.86 (m, 4H), 0.83 (m, 3H), 0.72 (m, 3H). MS(ESI) m/z 875.91 [M+H]⁺.

Example AM

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylic acidand(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(2S)-2-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)acetic acid. ¹H NMR (400 MHz, dmso) δ 8.64 (s, 1H), 8.29-7.49 (m, 7H),5.26 (s, 2H), 5.15 (m, 1H), 5.07-4.91 (m, 1H), 4.61 (m, 2H), 4.17-3.29(m, 16H), 2.43-2.02 (m, 8H), 1.83 (s, 2H), 1.47 (m, 5H), 1.36-0.76 (m,12H), 0.71 (m, 3H). MS (ESI) m/z 889.60 [M+H]⁺.

Example AN

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2R)-2-[(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2R)-2-[(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylic acidand(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(2R)-2-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)acetic acid. ¹H NMR (400 MHz, dmso) δ 8.59 (m, 2H), 8.27-7.25 (m, 6H),5.23 (m, 2H), 5.06-4.86 (m, 1H), 4.76-4.21 (m, 3H), 4.12-2.96 (m, 18H),2.51-1.69 (m, 12H), 1.65-1.33 (m, 6H), 1.25-0.55 (m, 8H), 0.07 (m, 2H).MS (ESI) m/z 889.69 [M+H]⁺.

Example AO

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid;(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid and(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(2S)-2-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)acetic acid ¹H NMR (400 MHz, dmso) δ 8.60 (s, 2H), 8.25-7.37 (m,8H), 5.22 (s, 2H), 5.11 (s, 1H), 4.95 (s, 1H), 4.67 (s, 1H), 4.52 (m,2H), 3.56 (m, 15H), 2.25 (m, 8H), 1.80 (s, 2H), 1.44 (m, 6H), 1.26-0.54(m, 12H) MS (ESI) m/z 889.56 [M+H]⁺.

Example AP

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2R)-2-[(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2R)-2-[(2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid;(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid and(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(2R)-2-((2R,6S)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid. ¹H NMR (400 MHz, dmso) δ 8.60 (m, 1H), 8.24-7.41 (m, 8H), 5.23 (m,2H), 5.10 (m, 2H), 4.65 (s, 2H), 3.73 (m, 14H), 2.33 (m, 11H), 1.84 (s,3H), 1.54-1.30 (m, 5H), 1.29-0.61 (m, 11H), 0.48 (s, 1H). MS (ESI) m/z890.05 [M+H]⁺.

Example AQ

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylic acid.¹H NMR (400 MHz, dmso) δ 8.63 (s, 1H), 8.24-7.44 (m, 8H), 5.25 (s, 2H),5.14 (s, 1H), 4.99 (s, 1H), 4.67 (m, 2H), 3.96 (m, 5H), 3.48 (m, 12H),2.44-1.75 (m, 9H), 1.48 (m, 6H), 1.30-1.10 (m, 3H), 1.01 (m, 3H), 0.85(m, 4H), 0.75 (m, 3H). MS (ESI) m/z 889.58 [M+H]⁺.

Example AR

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2R)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2R)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with (2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylic acidand(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(R)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)acetic acid. ¹H NMR (400 MHz, dmso) δ 8.64 (m, 1H), 7.67 (m, 7H),5.36-5.12 (m, 2H), 4.99 (s, 1H), 4.62 (s, 1H), 4.38 (s, 1H), 4.22 (s,1H), 4.16-4.02 (m, 1H), 4.00-3.84 (m, 1H), 3.70-3.09 (m, 12H), 2.24 (m,5H), 1.84 (s, 2H), 1.60 (s, 1H), 1.44 (m, 4H), 1.33-0.36 (m, 19H). MS(ESI) m/z 889.76 [M+H]⁺.

Example AS

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylic acid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidand (2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid. ¹H NMR (400 MHz, dmso) δ 8.65 (s, 1H), 8.23-7.44 (m, 8H), 5.26 (s,2H), 5.15 (s, 1H), 4.99 (s, 1H), 4.66 (m, 2H), 4.24-3.82 (m, 4H),3.75-3.20 (m, 12H), 2.42-1.72 (m, 10H), 1.47 (m, 5H), 1.30-0.96 (m, 6H),0.95-0.62 (m, 8H). MS (ESI) m/z 889.88 [M+H]⁺.

Example AT

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2R)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2R)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid;(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid and(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid with(R)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)acetic acid ¹H NMR (400 MHz, dmso) δ 8.65 (s, 1H), 7.74 (m, 8H),5.71-5.53 (m, 1H), 5.28 (s, 2H), 5.15 (s, 1H), 5.05 (s, 1H), 4.70 (s,1H), 4.13 (m, 5H), 3.82-3.15 (m, 10H), 2.70-2.57 (m, 1H), 2.43-1.71 (m,9H), 1.67-1.29 (m, 6H), 1.28-0.54 (m, 14H). MS (ESI) ink 889.53 [M+H]⁺.

Example AU

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamate

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid.¹H NMR (400 MHz, dmso) δ 12.94 (s, 1H), 12.36 (s, 1H), 11.77 (s, 1H),8.42 (m, 1H), 8.13-7.16 (m, 7H), 5.11 (s, 3H), 4.96 (s, 1H), 4.64 (s,2H), 3.97 (m, 4H), 3.67-3.11 (m, 13H), 2.39-1.66 (m, 10H), 1.62-1.30 (m,7H), 1.30-0.92 (m, 9H), 0.81 (m, 6H). MS (ESI) m/z 960.04 [M+H]⁺.

Example AV

methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid, and substituting(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith (2S,4S)-1-(tert-butoxycarbonyl)-4-methylpyrrolidine-2-carboxylicacid. ¹H NMR (400 MHz, Methanol-d₄) δ 8.63 (d, 1H), 8.19 (d, 1H),8.11-7.76 (m, 4H), 7.72-7.57 (m, 1H), 5.44-5.26 (m, 2H), 5.23-5.11 (m,1H), 5.00-4.71 (m, 5H), 4.47 (t, 1H), 4.16 (dt, 3H), 3.81-3.62 (m, 5H),3.53 (t, 1H), 2.83-2.67 (m, 1H), 2.53 (dd, 2H), 2.33 (dd, 2H), 2.04 (dd,4H), 1.54 (dd, 2H), 1.52-1.38 (m, 3H), 1.28 (d, 3H), 1.20 (s, 1H), 1.15(s, 2H), 1.11-0.95 (m, 6H), 0.87 (t, 2H). LCMS-ESI+calc'd forC49H61N8O8:890.05; Found: 889.23.

Example AW

methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid, and substituting(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith (S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid. 1H NMR(400 MHz, Methanol-d4) δ 8.65 (s, 1H), 8.21 (d, 1H), 8.08-7.96 (m, 1H),7.92 (s, 1H), 7.85 (d, 1H), 7.75-7.59 (m, 2H), 5.45-5.38 (m, 1H), 5.33(s, 1H), 5.23-5.11 (m, 1H), 4.35-4.06 (m, 4H), 4.01-3.92 (m, 1H),3.82-3.44 (m, 7H), 2.75-2.48 (m, 3H), 2.46-2.09 (m, 6H), 2.07-1.91 (m,2H), 1.56 (d, 3H), 1.49-1.36 (m, 2H), 1.32-1.21 (m, 2H), 1.15 (d, 3H),1.10-0.93 (m, 6H), 0.88 (d, 2H). LCMS-ESI+calc'd for C48H59N8O8:875.45;Found: 875.29.

Example AX

methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(5-{2-[(2S,3aS,6aS)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}octahydrocyclopenta[b]pyrrol-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid, and substituting(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith(2S,3aS,6a5)-1-(tert-butoxycarbonyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid. 1H NMR (400 MHz, Methanol-d4) δ 8.66 (s, 1H), 8.21 (d, 1H),8.12-7.97 (m, 2H), 7.94 (s, 1H), 7.91-7.78 (m, 1H), 7.74-7.62 (m, 2H),5.81-5.71 (m, 1H), 5.39-5.26 (m, 3H), 5.23-5.11 (m, 1H), 4.84-4.76 (m,2H), 4.25 (d, 1H), 4.21-4.06 (m, 2H), 3.81 (s, 1H), 3.74 (s, 1H), 3.67(d, 4H), 3.63-3.52 (m, 1H), 3.16-3.04 (m, 1H), 2.76-2.64 (m, 1H),2.61-2.48 (m, 1H), 2.44-2.29 (m, 3H), 2.23 (q, 2H), 2.18-2.08 (m, 2H),2.07-1.85 (m, 4H), 1.84-1.66 (m, 2H), 1.57 (d, 3H), 1.49-1.36 (m, 2H),1.29 (t, 1H), 1.27-1.18 (m, 1H), 1.15 (d, 3H), 1.12-1.01 (m, 4H), 0.98(d, 3H), 0.88 (d, 2H). LCMS-ESI+calc'd for C51H63N8O8:915.48; Found:915.29.

Example AY

methyl{(2S)-1-[(2S,3aS,6aS)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)hexahydrocyclopenta[b]pyrrol-1(2H)-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,3aS,6aS)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)hexahydrocyclopenta[b]pyrrol-1(2H)-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,3aS,6aS)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid. 1H NMR (400 MHz, Methanol-d4) δ 8.66 (s, 1H), 8.29-8.18 (m, 1H),8.10-8.03 (m, 1H), 7.94 (d, 1H), 7.84 (d, 1H), 7.73-7.64 (m, 1H), 5.32(d, 2H), 5.24-5.13 (m, 1H), 4.18 (dd, 2H), 3.86-3.72 (m, 2H), 3.67 (d,3H), 3.52 (d, 1H), 3.11-2.97 (m, 1H), 2.72-2.57 (m, 2H), 2.54-2.21 (m,3H), 2.20-1.80 (m, 7H), 1.79-1.70 (m, 1H), 1.65 (d, 2H), 1.61-1.52 (m,1H), 1.50-1.36 (m, 1H), 1.36-1.23 (m, 2H), 1.14 (d, 3H), 1.10-1.02 (m,1H), 1.02-0.88 (m, 6H). LCMS-ESI+calc'd for C51H63N8O8:915.48; Found:915.379.

Examples AZ, BA, BB, BC

methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,3S,5S)-5-(5-{2-[(2S,4S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4,5-dimethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-2,3-dimethylpyrrolidin-1-yl]-2-oxoethyl}carbamatemethyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2R,3R,5R)-5-(9-{2-[(2S,4S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4,5-dimethylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-2,3-dimethylpyrrolidin-1-yl]-2-oxoethyl}carbamatemethyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2R,3R,5R)-5-(5-{2-[(2S,4S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4,5-dimethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-2,3-dimethylpyrrolidin-1-yl]-2-oxoethyl}carbamatemethyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2R,3R,5R)-5-(5-{2-[(2R,4R,5R)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4,5-dimethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-2,3-dimethylpyrrolidin-1-yl]-2-oxoethyl}carbamate

Following Example AA, substitutingrel-(2S,4S,5S)-1-(tert-butoxycarbonyl)-4,5-dimethylpyrrolidine-2-carboxylicacid for(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid and(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acid,and using two equivalents of(S)-2-((2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl)-2-(methoxycarbonylamino)aceticacid, provided a mixture of four diastereomers. The diastereomers wereseparated by reverse phase HPLC (Gemini column, 10-45% MeCN/H2O/0.1%TFA).

Example AZ

Methyl{(1S)-1-{(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,3S,5S)-5-(5-{2-[(2S,4S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4,5-dimethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-2,3-dimethylpyrrolidin-1-yl]-2-oxoethyl}carbamate.RT=3.757 min (Gemini column, 2-95% MeCN/H2O/0.1% TFA over 8 min).LCMS-ESI+: calc'd for C55H70N8O9:986.53 (M+); Found: 987.88 (M+H+). ¹HNMR (400 MHz, Methanol-d₄) δ 8.25 (s, 1H), 7.96-7.78 (m, 1H), 7.76-7.34(m, 8H), 7.27 (s, 1H), 5.22-5.06 (m, 4H), 5.00 (t, 1H), 4.65-4.44 (m,2H), 4.17-3.95 (m, 4H), 3.57 (s, 6H), 3.55-3.46 (m, 1H), 3.45-3.33 (m,1H), 2.50-1.97 (m, 9H), 1.55 (t, 2H), 1.41 (d, 3H), 1.37-1.26 (m, 5H),1.24-1.13 (m, 1H), 1.13-1.01 (m, 12H), 1.01-0.93 (m, 1H), 0.86 (d, 3H),0.83-0.74 (m, 5H).

Example BA

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2R,3R,5R)-5-(9-{2-[(2S,4S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4,5-dimethylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-2,3-dimethylpyrrolidin-1-yl]-2-oxoethyl}carbamate.RT=3.899 min (Gemini column, 2-95% MeCN/H2O/0.1% TFA over 8 min).LCMS-ESI+: calc'd for C55H70N8O9:986.53 (M+); Found: 987.95 (M+H+). ¹HNMR (400 MHz, Methanol-d4) δ 8.43-8.28 (m, 1H), 8.05-7.88 (m, 1H),7.88-7.42 (m, 6H), 7.39-7.25 (m, 1H), 5.31-5.01 (m, 4H), 4.70-4.55 (m,1H), 4.46-4.18 (m, 2H), 4.18-4.05 (m, 2H), 4.04-3.94 (m, OH), 3.85-3.71(m, 1H), 3.65 (s, 5H), 3.52-3.38 (m, 1H), 2.63-1.98 (m, 9H), 1.75-1.50(m, 3H), 1.47 (d, 3H), 1.44-1.21 (m, 6H), 1.18-0.99 (m, 16H), 0.96-0.87(m, 4H), 0.87-0.82 (m, 3H), 0.65 (d, J=6.1 Hz, 1H).

Example BB

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2R,3R,5R)-5-(5-{2-[(2S,4S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4,5-dimethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-2,3-dimethylpyrrolidin-1-yl]-2-oxoethyl}carbamate.RT=3.940 min (Gemini column, 2-95% MeCN/H2O/0.1% TFA over 8 min).LCMS-ESI+: calc'd for C55H70N8O9:986.53 (M+); Found: 987.86 (M+H+). ¹HNMR (400 MHz, Methanol-d₄) δ 8.23-8.03 (m, 1H), 7.79-7.64 (m, 2H),7.59-7.20 (m, 6H), 7.15-7.02 (m, 1H), 5.04-4.87 (m, 3H), 4.83 (t, 1H),4.43-4.27 (m, 1H), 4.27-3.76 (m, 5H), 3.63-3.46 (m, 3H), 3.45-3.36 (m,5H), 2.55-1.68 (m, 10H), 1.56-1.27 (m, 3H), 1.23 (d, J=6.8 Hz, 1H),1.20-1.11 (m, 5H), 1.08 (t, J=7.1 Hz, 1H), 1.04-0.73 (m, 18H), 0.73-0.51(m, 5H), 0.01 (d, J=6.0 Hz, 1H).

Example BC

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2R,3R,5R)-5-(5-{2-[(2R,4R,5R)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4,5-dimethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-2,3-dimethylpyrrolidin-1-yl]-2-oxoethyl}carbamate.RT=4.076 min (Gemini column, 2-95% MeCN/H2O/0.1% TFA over 8 min).LCMS-ESI+: calc'd for C55H70N8O9: 986.53 (M+); Found: 987.91 (M+H+). ¹HNMR (400 MHz, Methanol-d₄) δ 8.22-8.08 (m, 1H), 7.81-7.67 (m, 1H),7.65-7.17 (m, 5H), 7.10 (s, 1H), 5.19-5.05 (m, 1H), 5.04-4.80 (m, 3H),4.30-3.90 (m, 6H), 3.63-3.46 (m, 4H), 3.40 (s, 5H), 2.55-1.60 (m, 10H),1.55-1.25 (m, 4H), 1.21-0.95 (m, 12H), 0.95-0.74 (m, 14H), 0.56 (d, 2H),0.49-0.34 (m, 1H), 0.05-0.04 (m, 1H).

Example BD

methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4-methoxymethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamate

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(5-{2-[(2S,4S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4-methoxymethylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidand (2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid ¹H NMR (400 MHz, dmso) δ 8.71 (s, 1H), 8.18 (m, 1H), 7.95 (m, 4H),7.80-7.54 (m, 3H), 7.45 (m, 1H), 5.34-5.14 (m, 3H), 5.05-4.92 (m, 1H),4.62 (s, 1H), 4.35-3.03 (m, 13H), 2.66 (s, 2H), 2.50 (m, 2H), 2.39-1.72(m, 9H), 1.55-0.67 (m, 28H). MS (ESI) ink 989.41 [M+H]⁺.

Example BE

methyl{(2S,3S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate

Methyl{(2S,3S)-1-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamatewas synthesized in a similar manner as example AA omitting the oxidationwith MnO₂. ¹H NMR (400 MHz, dmso) δ 7.84 (m, 4H), 7.62 (m, 2H), 7.53 (m,1H), 7.25 (s, 1H), 5.15 (s, 2H), 5.02-4.88 (m, 2H), 4.62 (s, 2H),4.14-3.24 (m, 16H), 3.06 (s, 2H), 2.88 (s, 2H), 2.21 (m, 8H), 1.82 (s,2H), 1.67 (s, 1H), 1.44 (m, 8H), 1.30-0.95 (m, 8H), 0.91 (m, 3H), 0.78(m, 5H), 0.64 (m, 3H). MS (ESI) m/z 905.78 [M+H]⁺.

Example BF

methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamate.

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,4,5,11-tetrahydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,4S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidand omitting the oxidation with MnO₂. ¹H NMR (400 MHz, dmso) δ 8.05-7.75(m, 4H), 7.73-7.47 (m, 3H), 7.31 (m, 1H), 5.15 (s, 2H), 4.96 (m, 2H),4.61 (s, 2H), 4.15-3.15 (m, 18H), 3.06 (s, 2H), 2.99-2.75 (m, 3H), 2.17(m, 8H), 1.82 (m, 2H), 1.53-1.32 (m, 6H), 1.30-1.17 (m, 2H), 1.14-0.87(m, 11H), 0.85-0.69 (m, 2H). MS (ESI) m/z 961.54 [M+H]⁺.

Example BG

methyl{(2S,3S)-1-[(2S,3aS,6aS)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)hexahydrocyclopenta[b]pyrrol-1(2H)-yl]-3-methyl-1-oxopentan-2-yl}carbamate

Methyl{(2S,3S)-1-[(2S,3aS,6aS)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)hexahydrocyclopenta[b]pyrrol-1(2H)-yl]-3-methyl-1-oxopentan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,3aS,6aS)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)octahydrocyclopenta[b]pyrrole-2-carboxylicacid. ¹H NMR (400 MHz, dmso) δ 8.62 (s, 1H), 8.18 (s, 2H), 7.90-7.64 (m,3H), 7.63-7.39 (m, 2H), 5.24 (s, 2H), 5.13 (s, 1H), 5.02 (s, 1H), 4.69(s, 2H), 4.13-3.85 (m, 3H), 3.47 (m, 10H), 2.87 (s, 2H), 2.42 (m, 2H),2.10 (s, 5H), 1.75 (m, 5H), 1.51 (m, 7H), 1.31-0.94 (m, 7H), 0.92-0.64(m, 9H). MS (ESI) m/z 929.46 [M+H]⁺.

Example BH

methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamate

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(9-{2-[(2S,4S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-4-(methoxymethyl)pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,4S)-1-(tert-butoxycarbonyl)-4-(methoxymethyl)pyrrolidine-2-carboxylicacid. ¹H NMR (400 MHz, cdcl₃) δ 8.69 (s, 1H), 8.28-8.09 (m, 2H),8.00-7.75 (m, 3H), 7.62 m, 2H), 7.50 (m, 1H), 5.27 (s, 2H), 5.14 (1H),4.73 (s, 1H), 4.33-3.25 (m, 20H), 2.66 (s, 1H), 2.58-2.28 (m, 8H),2.25-1.79 (m, 4H), 1.65-0.66 (m, 21H). MS (ESI) m/z 989.65 [M+H]⁺.

Example BI

methyl{(2S)-1-[(2S,3aS,6aS)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)hexahydrocyclopenta[b]pyrrol-1(2H)-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,3aS,6aS)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)hexahydrocyclopenta[b]pyrrol-1(2H)-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidand (2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylicacid with (2S,3aS,6a5)-1-((S)-2-(methoxycarbonylamino)-3methylbutanoyl)octahydrocyclopenta [b]pyrrole-2-carboxylic acid. ¹H NMR(400 MHz, dmso) δ 8.67 (s, 1H), 7.72 (m, 8H), 5.33-5.17 (m, 3H), 5.00(m, 1H), 4.77 (s, 1H), 4.62 (s, 1H), 4.17-3.86 (m, 5H), 3.49 (m, 10H),2.89 (s, 1H), 2.56-1.70 (m, 7H), 1.47 (m, 6H), 1.30-0.97 (m, 6H), 0.90(s, 4H), 0.81 (m, 8H). MS (ESI) m/z 915.37 [M+H]⁺.

Example BJ

methyl{(2S)-1-[(2S,5S)-2-(9-{2-[(2S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamate

Methyl{(2S)-1-[(2S,5S)-2-(9-{2-[2S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}pyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxobutan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidand (2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((S)-2-(methoxycarbonylamino)-3-methylbutanoyl)-5-methylpyrrolidine-2-carboxylicacid ¹H NMR (400 MHz, dmso) δ 8.67 (s, 1H), 8.19 (m, 1H), 8.03 (m, 2H),7.91-7.68 (m, 3H), 7.68-7.38 (m, 3H), 5.26 (s, 2H), 5.14 (m, 2H), 4.70(s, 1H), 4.20-3.23 (m, 14H), 2.37 (s, 2H), 2.22-1.71 (m, 6H), 1.49 (m,3H), 1.41-0.97 (m, 7H), 0.97-0.78 (m, 8H), 0.72 (m, 3H). MS (ESI) m/z875.30 [M+H]⁺.

Example BK

methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S)-2-(5-{2-[(2S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxoethyl}carbamate

Methyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S)-2-(5-{2-[(2S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}pyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)pyrrolidin-1-yl]-2-oxoethyl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with (S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid and(2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidwith (S)-1-(tert-butoxycarbonyl)pyrrolidine-2-carboxylic acid. ¹H NMR(400 MHz, dmso) δ 8.69 (s, 1H), 8.19 (m, 1H), 8.08 (s, 1H), 7.86 (m,3H), 7.73 (s, 1H), 7.62 (m, 1H), 7.47 (m, 1H), 5.26 (s, 3H), 5.10 (m,1H), 4.12-3.24 (m, 18H), 2.37 (s, 2H), 2.31-1.89 (m, 8H), 1.31 (m, 6H),1.06 (m, 7H), 0.96-0.76 (m, 8H). MS (ESI) m/z 931.86 [M+H]⁺.

Example BL

methyl{(2S,3S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamate

Methyl{(2S,3S)-1-[(2S,5S)-2-(9-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1H-imidazol-5-yl}-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-2-yl)-5-methylpyrrolidin-1-yl]-3-methyl-1-oxopentan-2-yl}carbamatewas synthesized in a similar manner as example AA substituting(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid with((2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylic acidand (2S,5S)-1-(tert-butoxycarbonyl)-5-methylpyrrolidine-2-carboxylicacid with(2S,5S)-1-((2S,3S)-2-(methoxycarbonylamino)-3-methylpentanoyl)-5-methylpyrrolidine-2-carboxylicacid. ¹H NMR (400 MHz, dmso) δ 8.66 (s, 1H), 8.18 (m, 1H), 8.12-7.71 (m,5H), 7.58 (m, 2H), 5.27 (s, 2H), 5.15 (m, 1H), 5.06-4.93 (m, 1H), 4.67(m, 2H), 4.22-3.29 (m, 11H), 2.23 (m, 7H), 1.83 (s, 2H), 1.65 (s, 1H),1.47 (m, 8H), 1.29-0.98 (m, 7H), 0.95-0.70 (m, 8H), 0.66 (m, 3H). MS(ESI) m/z 903.87 [M+H]⁺.

Example BM

Ethyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(ethoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamate

Iodotrimethylsilane (1.14 ml, 8.03 mmol) is added to a solution ofmethyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(methoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamate(770 mg, 0.8 mmol) in Dichloromethane (10 ml), and the mixture is thenrefluxed for 3 hours. After cooling to room temperature the mixture wasconcentrated; dissolved in ethyl acetate and extracted with 2×7 ml of 1Nhydrochloric acid solution. The aqueous phases are combined, cooled andthen basified by addition of 5N sodium hydroxide. The basic aqueousphase is extracted with ethyl acetate 3×10 ml. The organic phases werecombined, dried over Na₂SO₄ and concentrated under vacuum, the product667 mg (98.5%) was treated with sodium hydroxide (66.25 mg, 1.66 mmol)in Water (7 ml). The mixture was cooled in an ice bath and ethylchloroformate (0.16 ml, 1.66 mmol) was added, the reaction mixture wasstirred at 0° C. for 30 min, extracted with 2×10 ml ethyl acetate. Thecombined organic layers were dried over Na₂SO₄ and concentrated. Theresidue was purified by reverse phase HPLC (Gemini column, 10-46%MeCN/H₂O/0.1% TFA). The desired fractions were combined, lyophilized toprovide ethyl{(1S)-1-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(2S,5S)-2-(5-{2-[(2S,5S)-1-{(2S)-2-[(2R,6R)-2,6-dimethyltetrahydro-2H-pyran-4-yl]-2-[(ethoxycarbonyl)amino]acetyl}-5-methylpyrrolidin-2-yl]-1,11-dihydroisochromeno[4′,3′:6,7]naphtho[1,2-d]imidazol-9-yl}-1H-imidazol-2-yl)-5-methylpyrrolidin-1-yl]-2-oxoethyl}carbamate.¹H NMR (400 MHz, dmso) δ 8.62 (s, 1H), 8.26-7.33 (m, 9H), 5.24 (s, 2H),5.13 (s, 1H), 4.98 (s, 1H), 4.80-4.55 (m, 2H), 4.30-3.23 (m, 9H), 2.19(m, 9H), 1.83 (s, 2H), 1.48 (m, 10H), 1.32-0.69 (m, 24H). MS (ESI) m/z987.89 [M+H]⁺.

Biological Assays

Effect of Serum Proteins on Replicon Potency:

Replicon assays are conducted in normal cell culture medium (DMEM+10%FBS) supplemented with physiologic concentrations of human serum albumin(40 mg/mL) or α-acid glycoprotein (1 mg/mL). EC₅₀s in the presence ofhuman serum proteins are compared to the EC₅₀ in normal medium todetermine the fold shift in potency.

MT-4 Cell Cytotoxicity:

MT4 cells are treated with serial dilutions of compounds for a five dayperiod. Cell viability is measured at the end of the treatment periodusing the Promega CellTiter-Glo assay and non-linear regression isperformed to calculate CC₅₀.

Compound Concentration Associated with Cells at EC₅₀:

Huh-luc cultures are incubated with compound at concentrations equal toEC₅₀. At multiple time points (0-72 hours), cells are washed 2× withcold medium and extracted with 85% acetonitrile; a sample of the mediaat each time-point will also be extracted. Cell and media extracts areanalyzed by LC/MS/MS to determine the Molar concentration of compoundsin each fraction. Representative compounds of the disclosure have shownactivity.

Solubility and Stability:

Solubility is determined by taking an aliquot of 10 mM DMSO stocksolution and preparing the compound at a final concentration of 100 μMin the test media solutions (PBS, pH 7.4 and 0.1 N HCl, pH 1.5) with atotal DMSO concentration of 1%. The test media solutions are incubatedat room temperature with shaking for 1 hr. The solutions will then becentrifuged and the recovered supernatants are assayed on the HPLC/UV.Solubility will be calculated by comparing the amount of compounddetected in the defined test solution compared to the amount detected inDMSO at the same concentration. Stability of compounds after an 1 hourincubation with PBS at 37° C. will also be determined.

Stability in Cryopreserved Human, Dog, and Rat Hepatocytes:

Each compound is incubated for up to 1 hour in hepatocyte suspensions(100 μl, 80,000° Cells per well) at 37° C. Cryopreserved hepatocytes arereconstituted in the serum-free incubation medium. The suspension istransferred into 96-well plates (50 μL/well). The compounds are dilutedto 2 μM in incubation medium and then are added to hepatocytesuspensions to start the incubation. Samples are taken at 0, 10, 30 and60 minutes after the start of incubation and reaction will be quenchedwith a mixture consisting of 0.3% formic acid in 90% acetonitrile/10%water. The concentration of the compound in each sample is analyzedusing LC/MS/MS. The disappearance half-life of the compound inhepatocyte suspension is determined by fitting the concentration-timedata with a monophasic exponential equation. The data will also bescaled up to represent intrinsic hepatic clearance and/or total hepaticclearance.

Stability in Hepatic S9 Fraction from Human, Dog, and Rat:

Each compound is incubated for up to 1 hour in S9 suspension (500 μl, 3mg protein/mL) at 37° C. (n=3). The compounds are added to the S9suspension to start the incubation. Samples are taken at 0, 10, 30, and60 minutes after the start of incubation. The concentration of thecompound in each sample is analyzed using LC/MS/MS. The disappearancehalf-life of the compound in S9 suspension is determined by fitting theconcentration-time data with a monophasic exponential equation.

Caco-2 Permeability:

Compounds are assayed via a contract service (Absorption Systems, Exton,Pa.). Compounds are provided to the contractor in a blinded manner Bothforward (A-to-B) and reverse (B-to-A) permeability will be measured.Caco-2 monolayers are grown to confluence on collagen-coated,microporous, polycarbonate membranes in 12-well Costar TRANSWELL®plates. The compounds are dosed on the apical side for forwardpermeability (A-to-B), and are dosed on the basolateral side for reversepermeability (B-to-A). The cells are incubated at 37° C. with 5% CO₂ ina humidified incubator. At the beginning of incubation and at 1 hr and 2hr after incubation, a 200-μL aliquot is taken from the receiver chamberand replaced with fresh assay buffer. The concentration of the compoundin each sample is determined with LC/MS/MS. The apparent permeability,Papp, is calculated.

Plasma Protein Binding:

Plasma protein binding is measured by equilibrium dialysis. Eachcompound is spiked into blank plasma at a final concentration of 2 μM.The spiked plasma and phosphate buffer is placed into opposite sides ofthe assembled dialysis cells, which will then be rotated slowly in a 37°C. water bath. At the end of the incubation, the concentration of thecompound in plasma and phosphate buffer is determined. The percentunbound is calculated using the following equation:

${\% \mspace{14mu} {Unbound}} = {100\mspace{11mu} \bullet \mspace{11mu} \left( \frac{C_{f}}{C_{b} + C_{f}} \right)}$

Where C_(f) and C_(b) are free and bound concentrations determined asthe post-dialysis buffer and plasma concentrations, respectively.

Cyp450 Profiling:

Each compound is incubated with each of 5 recombinant human CYP450enzymes, including CYP1A2, CYP2C9, CYP3A4, CYP2D6 and CYP2C19 in thepresence and absence of NADPH. Serial samples will be taken from theincubation mixture at the beginning of the incubation and at 5, 15, 30,45 and 60 minutes after the start of the incubation. The concentrationof the compound in the incubation mixture is determined by LC/MS/MS. Thepercentage of the compound remaining after incubation at each time pointis calculated by comparing with the sampling at the start of incubation.

Stability in Rat, Dog, Monkey and Human Plasma:

Compounds will be incubated for up to 2 hours in plasma (rat, dog,monkey, or human) at 37° C. Compounds are added to the plasma at finalconcentrations of 1 and 10 μg/mL. Aliquots are taken at 0, 5, 15, 30,60, and 120 minutes after adding the compound. Concentration ofcompounds and major metabolites at each time point are measured byLC/MS/MS.

Evaluation of Cell-Based Anti-HCV Activity:

Antiviral potency (EC₅₀) was determined using a Renilla luciferase(RLuc)-based HCV replicon reporter assay. To perform the assay forgenotype 1 and 2a JFH-1, stable HCV 1a RLuc replicon cells (harboring adicistronic genotype 1a H77 replicon that encodes a RLuc reporter),stable HCV 1b RLuc replicon cells (harboring a dicistronic genotype 1bCon1 replicon that encodes a RLuc reporter), or stable HCV 2a JFH-1 Rlucreplicon cells (harboring a dicistronic genotype 2a JFH-1 replicon thatencodes a RLuc reporter; with L31 present in NS5A) were dispensed into384-well plates for EC₅₀ assays. To perform the assay for genotype 2a(with M31 present in NS5A) or 2b, NS5A chimeric genotype 2a JFH-1replicons that encodes a RLuc-Neo reporter and either genotype 2a J6strain NS5A gene or genotype 2b MD2b-1 strain NS5A gene (both with M31present) respectively, were either transiently transfected (t) intoHuh-Lunet cells or were established as stably replicating replicon cells(s) is provided. Either cells were dispensed into 384-well plates forEC₅₀ assays. To perform the assay for genotype 3 and 4, NS5A chimericgenotype 1b Con1 replicons that encodes a Pi-RLuc reporter and eithergenotype 3a S52 strain NS5A gene or genotype 4a ED43 strain NS5A generespectively, were transiently transfected (t) into Huh-Lunet cells,which were subsequently dispensed into 384-well plates. Compounds weredissolved in DMSO at a concentration of 10 mM and diluted in DMSO eithermanually or using an automated pipeting instrument. Serially 3-folddiluted compounds were either manually mixed with cell culture media andadded to the seeded cells or directly added to the cells using anautomated instrument. DMSO was used as a negative (solvent; noinhibition) control, and the protease inhibitor ITMN-191 was included ata concentration>100× EC₅₀ as a positive control. 72 hours later, cellswere lysed and Renilla luciferase activity quantified as recommended bythe manufacturer (Promega-Madison, Wis.). Non-linear regression wasperformed to calculate EC₅₀ values.

To determine the antiviral potency (EC₅₀) against resistance mutants,resistance mutations, including M28T, Q30R, Q30H, Q30E, L31M, Y93C,Y93H, and Y93N in genotype 1a NS5A, Y93H and L31V/Y93H in genotype 1bNS5A, and Y93H for in genotype 3a NS5A, were introduced individuallyinto either 1a Pi-Rluc or 1b Pi-Rluc replicons by site directedmutagenesis. Replicon RNA of each resistant mutant was transientlytransfected into Huh-7-derived cured-51 cells and antiviral potency wasdetermined on these transfected cells as described above.

IV and PO Single Dose Pharmacokinetic Studies in SD Rats:

The pharmacokinetics of selected compounds was characterized in maleSprague-Dawley (SD) rats (250-300g). In this study, two groups of naïvepurebred SD rats (N=3 per group, fasted over night) received theselected compound either as an intravenous (IV) infusion (1 mg/kg over30 minutes) via the jugular vein or by oral gavage (2 mg/kg). Theintravenous (IV) dosing vehicle was 5% ethanol, 35% polyethylene glycol400 (PEG 400) and 60% water pH 2.0. The oral dosing vehicle was 5%ethanol, 55% PEG 400 and 40% citrate buffer pH 2.2.

Serial blood samples (approximately 0.3 mL each) were collected fromjugular vein or other suitable vein at specified time points. For the IVinfusion group, the blood samples were collected predose and at 0.25,0.48, 0.58, 0.75, 1.5, 3, 6, 8, 12 and 24 hours after the start ofinfusion. For the oral group, the blood samples were collected predoseand at 0.25, 0.50, 1, 2, 4, 6, 8, 12 and 24 hours after dosing. Theblood samples were collected into Vacutainer™ tubes containing EDTA-K₃as the anti-coagulant and were centrifuged at approximately 4° C. toobtain plasma. The plasma samples were stored at −20° C. until analysisby LC/MS/MS.

A bioanalytical method utilizing high performance liquid chromatographycoupled to tandem mass spectrometry (LC/MS/MS) was developed foranalysis of the selected compound in rat plasma. Detection was performedusing selected reaction monitoring (SRM); Ions representing theprecursor (M+H)⁺ species was selected in quadrupole 1 (Q1) and collidedwith argon gas in the collision cell (Q2) to generate specific production, which was subsequently monitored by quadrupole 3 (Q3). Standardcurve and quality control samples were prepared in male rat plasma andprocessed in the same way as the test samples to generate quantitativedata.

Pharmacokinetic parameters were generated using non-compartmentalpharmacokinetic analysis (Phoenix WinNonlin, version 6.3). Values belowthe lower limit of quantification (LLOQ) were assigned a value of zeroif predose and treated as missing thereafter. Area under the curve (AUC)was calculated using the linear trapezoidal rule. The oralbioavailability (% F) was determined by comparison of the area under thecurve (AUC) of the compound and/or a metabolite generated in plasmafollowing oral administration to that generated following intravenousadministration.

Comparative examples (Comp 1-14) as shown in Tables 2A and 2B below wereprepared according to the synthetic protocols described herein using theappropriate starting materials.

TABLE 1 Ex. No. 2a 2a 1b L31V/ 1a 1a 1a 1a 3a 1b 1a JFH J6 (t) 2b (t) 2b(s) 3a 4a (s) Rat Y93H Q30R Q30E Y93H Y93N Y93H (nM) (nM) (nM) (nM) (nM)(nM) (nM) (nM) % F (nM) (nM) (nM) (nM) (nM) (nM) AE 0.022 0.024 0.0120.042 0.038 0.027 0.011 0.230 1.121 6.933 AF 0.037 0.022 0.012 0.0030.014 0.016 0.005 0.180 0.097 0.268 0.899 4.586 AU 0.039 0.029 0.0130.008 0.017 0.023 0.029 0.017 16.4 0.059 0.042 0.026 0.089 0.130 AC 0.030.025 0.016 0.003 0.020 0.019 0.005 1.332 0.312 0.832 1.634 14.495 AB0.046 0.026 0.011 0.028 0.024 0.025 0.006 0.668 0.082 0.163 0.418 8.769AA 0.017 0.016 0.011 0.006 0.014 0.029 0.014 0.008 28.1 0.498 0.1450.322 0.570 0.544 AT AR 0.09 8.983 2.106 5.137 15.804 8.304 4.191 AS0.045 0.026 0.028 0.015 0.026 0.064 0.051 0.024 17.4 2.033 0.115 0.1480.325 0.732 AQ 0.029 0.019 0.016 0.011 0.016 0.032 0.017 0.012 22.90.908 0.074 0.206 0.337 0.703 AP 0.038 1.518 1.101 4.444 4.444 4.4441.188 AO 0.015 0.012 0.008 0.017 0.015 0.045 0.021 0.018 50.3 17.4711.784 9.034 9.057 AN 0.049 4.444 0.852 4.444 4.444 4.444 0.935 AM 0.0140.009 0.005 0.008 0.008 0.023 0.011 0.01 21.9 11.641 1.213 4.651 8.060AJ 0.021 0.016 0.019 0.059 0.057 0.035 0.018 44.000 0.160 2.700 4.3033.732 AI 0.018 0.014 0.017 0.041 0.058 0.028 0.012 21.424 0.208 2.1064.054 2.838 AD 0.031 0.021 0.027 1.017 0.351 0.074 0.018 AH 0.021 0.0260.031 0.172 0.104 0.154 0.017 AG 0.026 0.023 0.025 0.121 0.092 0.1370.014 27.8 AL 0.03 0.02 0.026 0.776 0.713 0.086 0.028 BC 2.474 10.1388.039 23.058 23.3 6.235 BB 0.205 3.855 2.958 10.555 10.35 2.537 20.22329.566 BA 0.061 0.169 0.086 0.202 0.231 0.108 0.359 0.496 40.645 AZ0.031 0.021 0.01 0.024 0.025 0.013 0.025 0.059 4.516 BD 0.096 0.06 0.0170.026 0.072 0.029 0.088 0.104 0.814 BE 0.019 0.017 0.011 0.029 0.0220.011 0.755 1.423 2.253 BF 0.049 0.048 0.021 0.035 0.042 0.065 0.0250.035 0.125 0.365 BG 0.018 0.02 0.017 0.045 0.039 0.028 0.017 0.3640.883 5.583 AX 0.022 0.017 0.012 0.046 0.027 0.015 0.01 0.480 0.8854.966 BH 0.118 0.053 0.013 0.055 0.027 0.056 0.024 2.7 0.040 0.082 0.612BI 0.024 0.012 0.011 0.022 0.033 0.017 0.011 0.184 0.466 3.451 AY 0.0280.027 0.018 0.013 0.030 0.023 0.011 0.531 1.180 8.484 AV 0.036 0.0360.011 0.008 0.015 0.029 0.013 0.140 1.053 2.388 BJ 0.022 0.027 0.0090.066 0.029 0.025 0.011 0.506 1.490 1.269 AW 0.07 0.029 0.011 0.0210.029 0.015 0.008 0.704 1.473 1.506 BK 0.496 0.171 0.02 0.033 0.048 0.080.021 3.2 0.100 0.251 0.263 BL 0.024 0.018 0.009 0.022 0.021 0.014 0.0060.228 0.531 1.047 BM 0.053 0.034 0.023 0.143 0.039

TABLE 2A Ex- 2a 2a 2b 2b ample 1b 1a JFH J6 (t) (t) (s) 3a Structure No.(nM) (nM) (nM) (nM) (nM) (nM) (nM)

Comp 1 0.027 0.018 0.008 0.012 0.112 0.042

Comp 2 0.064 0.055 0.025 0.083 0.193 0.126

Comp 3 0.026 0.021 0.019 0.061 0.038 0.095 0.065

Comp 4 0.028 0.023 0.018 0.026 0.044 0.074 0.043

Comp 5 0.029 0.022 0.013 0.012 0.024 0.054 0.041

Comp 6 0.051 0.024 0.012 0.012 0.092 0.038 0.075

Comp 7 0.028 0.018 0.015 0.035 0.037 0.088 0.035

Comp 8 0.04 0.028 0.012 0.113 0.071

Comp 9 0.034 0.028 0.019 0.025 0.143 0.083

Comp 10 0.024 0.015 0.021 0.024 0.033 0.062 0.052

Comp 11 0.416 0.203 0.041 0.2 0.203 0.447

Comp 12 0.022 0.013 0.009 0.032 0.07 0.026

Comp 13 0.048 0.025 0.033 0.022 0.048 0.077

Comp 14 0.025 0.025 0.01 0.053 0.036

TABLE 2B 1b L31V/ 1a 1a 1a 1a 3a Example 4a (s) Rat Y93H Q30R Q30E Y93HY93N Y93H Structure No. (nM) % F (nM) (nM) (nM) (nM) (nM) (nM)

Comp 1 0.016 9.9 0.971 0.061 0.319 2.047 8.073

Comp 2 0.022

Comp 3 0.014 46.6 0.052 0.526 2.537 14.074

Comp 4 0.021 22.1 0.032 0.432 0.978 1.062 0.695

Comp 5 0.021 22 0.031 0.297 0.466 0.549

Comp 6 0.014 0.042 0.101 0.442 0.252

Comp 7 0.019 10.2

Comp 8 0.979 1.901 4.576

Comp 9 40.9 0.571 0.260 3.495 10.580 40.9

Comp 10 21.8 0.024 0.123 0.716 3.739 21.8

Comp 11

Comp 12 17.7 5.643 0.097 1.283 1.244 1.379 17.7

Comp 13 4.4 0.371 1.074 2.128 4.4

Comp 14 6.882 0.130 1.195 3.664 1.746

1. A compound of formula (I):E^(1a)-V^(1a)—C(═O)—P^(1a)—W^(1a)—P^(1b)—C(═O)—V^(1b)-E^(1b)  (I)wherein: W^(1a) is

and W^(1a) is optionally substituted with one or more halo, alkyl,haloalkyl, optionally substituted aryl, optionally substitutedheterocycle, or cyano; Y⁵ is —O—CH₂—, —CH₂—O—, —O—C(═O)—, or —C(═O)—O—;X⁵ is —CH₂—CH₂—, or —CH═CH—; P^(1a) and P^(1b) are each independently:

V^(1a) and V^(1b) are each independently:

provided that at least one of V^(1a) and V^(1b) is

E^(1a) and E^(1b) are each independently —N(H)(alkoxycarbonyl),—N(H)(cycloalkylcarbonyl), or —N(H)(cycloalkyloxycarbonyl); orE^(1a)-V^(1a) taken together are R^(9a); or E^(1b)-V^(1b) taken togetherare R^(9b); and R^(9a) and R^(9b) are each independently:

or a pharmaceutically acceptable salt or prodrug thereof.
 2. Thecompound of claim 1, which has the formula:

wherein the imidazole ring shown in formula A2 and A4 is optionallysubstituted with one or more halo, haloalkyl, cyano, or alkyl; or apharmaceutically acceptable salt or prodrug thereof.
 3. The compound ofclaim 1, wherein P^(1a) and P^(1b) are each independently:


4. The compound of claim 1, wherein: W^(1a) is

and W^(1a) is optionally substituted with one or more halo, alkyl,haloalkyl, or cyano; Y⁵ is —O—CH₂—, or —CH₂—O—; X⁵ is —CH₂—CH₂—, or—CH═CH—; P^(1a) and P^(1b) are each independently:

V^(1a) and V^(1b) are each independently:

provided that at least one of V^(1a) and V^(1b) is

E^(1a) and E^(1b) are each independently —N(H)(alkoxycarbonyl),—N(H)(cycloalkylcarbonyl), or —N(H)(cycloalkyloxycarbonyl); orE^(1a)-V^(1a) taken together are R^(9a); or E^(1b)-V^(1b) taken togetherare R^(9b); and R^(9a) and R^(9b) are each independently:

or a pharmaceutically acceptable salt or prodrug thereof.
 5. Thecompound of claim 1, wherein one of V^(1a) and V^(1b) is:


6. The compound of claim 1, wherein both of V^(1a) and V^(1b) are:


7. The compound of claim 1, wherein one of V^(1a) and V^(1b) is:


8. The compound of claim 1, wherein both of V^(1a) and V^(1b) are:


9. The compound of claim 1, wherein one of P^(1a) and P^(1b) is:


10. The compound of claim 1, wherein one of P^(1a) and P^(1b) is:


11. The compound of claim 1, wherein both of P^(1a) and P^(1b) are:


12. The compound of claim 1 wherein —V^(1a)—C(═O)—P^(1a)— and—P^(1b)—C(═O)—V^(1b)-are each independently:

provided that at least one of V^(1a) and V^(1b) is


13. The compound of claim 1 selected from the group consisting of:

or a pharmaceutically acceptable salt or prodrug thereof.
 14. Thecompound of claim 1 selected from the group consisting of:

or a pharmaceutically acceptable salt or prodrug thereof.
 15. Apharmaceutical composition comprising the compound as described in claim1 or a pharmaceutically acceptable salt, or prodrug thereof; and atleast one pharmaceutically acceptable carrier.
 16. The pharmaceuticalcomposition of claim 15, further comprising at least one additionaltherapeutic agent for treating HCV.
 17. The pharmaceutical compositionof claim 16, wherein said at least one additional therapeutic agent isselected from ribavirin, an NS3 protease inhibitor, a nucleoside ornucleotide inhibitor of HCV NS5B polymerase, an alpha-glucosidase 1inhibitor, a hepatoprotectant, a non-nucleoside inhibitor of HCVpolymerase, and combinations thereof.
 18. A method of treating hepatitisC, said method comprising administering to a human patient apharmaceutical composition which comprises a therapeutically effectiveamount of the compound as described in claim 1 or a pharmaceuticallyacceptable salt, or prodrug thereof.
 19. The method according to claim18, further comprising administering at least one additional fortreating HCV to the patient.
 20. The method according to claim 19,wherein said at least one additional therapeutic agent is selected fromribavirin, an NS3 protease inhibitor, a nucleoside or nucleotideinhibitor of HCV NS5B polymerase, an alpha-glucosidase 1 inhibitor, ahepatoprotectant, a non-nucleoside inhibitor of HCV polymerase, andcombinations thereof.